OBJECTIVE To provide a scientific basis for the quality evaluation and clinical application of Qingwen hufei granules. METHODS Fourteen batches of Qingwen hufei granules were used as samples to establish high-performance liquid chromatography （HPLC） fingerprints using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 Edition）. The chromatographic peaks were identified and the similarity was evaluated. Cluster analysis （CA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to conduct chemical pattern recognition analysis on the 14 batches of samples. Meanwhile， the contents of neochlorogenic acid （NGA）， chlorogenic acid （CHA）， cryptochlorogenic acid （CGA）， forsythoside A （FTA）， 3，5-O-dicaffeoylquinic acid （3，5-O- DA）， 4，5-O-dicaffeoylquinic acid （4，5-O-DA）， and angoroside C （AGC） in the samples were determined by HPLC. RESULTS The methodological investigation results of both the fingerprint and the content determination complied with the relevant requirements. Fourteen common peaks were indicated in the HPLC fingerprints of the 14 batches of samples， and 7 of them were identified [NGA （peak 2）， CHA （peak 3）， CGA （peak 5）， FTA （peak 11）， 3，5-O-DA （peak 12）， 4，5-O-DA （peak 13）， and AGC （peak 14）]； the similarity of each sample was greater than 0.94. The results of CA and PCA showed that the samples could be classified into 3 categories； the results of OPLS-DA indicated that peak 4 （unknown）， peak 11 （FTA）， peak 8 （unknown）， peak 9 （unknown）， and peak 1 （unknown） were the differential components. The content ranges of NGA， CHA， CGA， 3，5-O-DA， FTA， 4，5-O-DA and AGC in the 14 batches of samples were 0.210 4-0.458 7， 0.269 1-0.506 3， 0.228 1-0.461 1， 0.443 9-1.044 6， 0.066 7-0.155 7， 0.062 8-0.143 8， and 0.057 4-0.105 7 mg/g， respectively. CONCLUSIONS The HPLC fingerprint and multi-component content determination methods established in this study are efficient and reliable， and can be used for the quality evaluation of Qingwen hufei granules.

Hepatocellular carcinoma (HCC) is one of the fastest growing cancers in the world, ranking fourth among the causes of cancer-induced death in the world. At present, the field of HCC treatment is developing rapidly, and immunotherapy has been recognized as a promising treatment method, in which T cells play a key role in HCC immunotherapy. However, in the case of virus infection or in tumor microenvironment (TME), T cells will be continuously stimulated by antigens and then fall into the state of T cell exhaustion (Tex). This state will not only reduce the immunity of patients but also lead to poor efficacy of immunotherapy. Therefore, to deeply analyze the mechanism of Tex and to explore effective strategies to reverse Tex is the key point in the immunotherapy for HCC. This review aims to summarize the mechanism of Tex in HCC patients, and the current situation and shortcomings of drug research and development to reverse Tex at this stage, in order to provide theoretical basis for the optimization of immunotherapy regimen for HCC patients.
Humans ; Carcinoma, Hepatocellular/therapy* ; Liver Neoplasms/therapy* ; Immunotherapy/methods* ; T-Lymphocytes/immunology* ; Tumor Microenvironment/immunology* ; Animals ; T-Cell Exhaustion

Objective:To investigate the main influencing factors of mild cognitive impairment(MCI)in the elderly and explore the interaction between high fluoride exposure and related co-factors on MCI.Methods:A cross-sectional study was conducted from May to December 2024 in four towns in the rural areas of Tianjin(two historically high-fluoride towns and two non-high-fluoride towns). A total of 125 elderly people aged 60 years and above were randomly selected from each township.MCI was diagnosed using the Mini-Mental State Examination(MMSE)and Montreal Cognitive Assessment-Basic(MoCA-B)according to the diagnostic criteria for MCI.Binary logistic regression was used to analyze the influencing factors of MCI, and conditional logistic regression was employed to assess multiplicative and additive interaction effects.Results:A total of 481 participants were included, with 354 in the normal group and 127 in the MCI group, and the incidence of MCI was 26.40%.Univariate analysis showed that MCI was associated with age, annual household income per capita, high fluoride exposure, life stress, depression, and abnormal urinalysis( χ2=4.241, 4.017, 4.552, 7.143, 4.151, 5.113, all P<0.05). Multivariate logistic regression revealed that after adjusting for other confounders, high fluoride exposure( OR=1.816, 95% CI: 1.177-2.802), age ≥70 years( OR=1.584, 95% CI: 1.034-2.428), depression( OR=2.106, 95% CI: 1.042-4.254), and abnormal urinalysis( OR=1.595, 95% CI: 1.041-2.444)increased the risk of MCI.Compared with severe life stress, moderate stress( OR=0.254, 95% CI: 0.082-0.790)reduced the risk of MCI.No multiplicative or additive interaction was found between high fluoride exposure and depression, life stress, or abnormal urinalysis. Conclusions:High fluoride exposure, age ≥70 years, depression, abnormal urinalysis, and severe life stress may increase the risk of MCI in the elderly, but there is no interaction among them.

Objective To explore the association between different levels of fluoride exposure and dyslipidemia in elderly people, and to analyze the influencing factors and their interactions. Methods A total of 1 143 elderly people over 60 years old were randomly selected from historical high water fluorosis areas and control areas in Tianjin. Logistic regression model and classification tree model were used to analyze the influencing factors of dyslipidemia, and to analyze the interaction between high fluoride exposure and relevant influencing factors on dyslipidemia. Results The prevalence of elevated low density lipoprotein cholesterol (LDL-C) was 5.69% (65/1 143). There was a significant difference in the prevalence of high LDL-C in different fluoride-exposed areas (2 = 0.092，P = 0.762). Multivariate logistic analysis showed that high fluoride exposure (OR=2.306，95%CI：1.185-4.491) and abdominal obesity (OR=2.274，95%CI：1.299-3.978) were risk factors for high LDL-C, while type B personality (OR=0.529，95%CI：0.308-0.908) was a protective factor for high LDL-C. The results of classification tree model showed that abdominal obesity contributed the most to the prevalence of high LDL-C in the elderly, followed by high fluoride exposure and hyperglycemia. There was a significant multiplicative interaction between high fluoride exposure and abdominal obesity on dyslipidemia (OR=5.191,95%CI：1.609-16.745，P=0.006). Conclusion High fluoride exposure may increase the risk of high LDL-C, and there is a multiplicative interaction between high fluoride exposure and abdominal obesity on dyslipidemia.

Objective: To understand the dose response relationship between urinary fluoride and dental fluorosis among children from drinking water borne endemic fluorosis areas in Tianjin, so as to provide a scientific basis for assessing fluoride exposure risk among children from endemic areas and establishing reference values for urinary fluoride. Methods: From January to December 2024, 83 endemic villages were selected in Tianjin. A total of 2 382 children aged 8-12 years from these villages underwent dental fluorosis examination, along with water fluoride and urinary fluoride testing. Additionally, data from areas where the drinking water fluorosis control target was achieved for more than 12 years (10 villages, 50 people per village) were included as controls. A restricted cubic spline model was used to analyze the relationship between urinary fluoride levels and dental fluorosis prevalence, and benchmark dose (BMD) and benchmark dose lower bound (BMDL), as well as reference dose (RfD), were calculated using the benchmark dose method. Results: The prevalence rates of dental fluorosis among children in drinking water borne endemic fluorosis areas and areas of normal fluoride content in water in Tianjin were 10.58%, 7.60%, with a geometric mean urinary fluoride level of 0.72 and 0.60 mg/L,respectively. There were statistically significant differences in both dental fluorosis prevalence and geometric mean urinary fluoride levels between the drinking water borne endemic fluorosis areas group and the areas of normal fluoride content in water ( χ 2/Z = 4.05 , -7.31, both P <0.05). Across different periods of water source improvement, there were statistically significant differences in overall population, male, and female dental fluorosis prevalence rates and geometric mean urinary fluoride levels ( χ 2/H =44.95, 23.96, 21.05; 168.39, 63.93, 107.50, all P <0.01). Significant differences were also observed across age groups among children from drinking water borne endemic fluorosis areas in terms of dental fluorosis prevalence and geometric mean urinary fluoride levels ( χ 2/H =32.14, 79.73, both P <0.01). The results of the restricted cubic spline model showed that the risk of dental fluorosis in different sex, age and overall children in the drinking water borne endemic fluorosis areas increased significantly with rising urinary fluoride concentration(all P-general trend <0.05). The BMD value for the drinking water borne endemic fluorosis areas was 1.72 mg/L, the BMDL was 1.31 mg/L, and the RfD was 1.31 mg/L. Conclusions The prevalence of dental fluorosis among children in the drinking water borne endemic fluorosis areas in Tianjin has been effectively controlled, but it is still higher than that in the areas of normal fluoride content in water. Moreover, urinary fluoride levels and dental fluorosis prevalence among children from drinking water borne endemic fluorosis areas show a clear dose response relationship.

BACKGROUND:Fibromyalgia syndrome,as a common rheumatic disease,is related to central sensitization and immune abnormalities.However,the specific mechanism has not been elucidated,and there is a lack of specific diagnostic markers.Exploring the possible pathogenesis of this disease has important clinical significance. OBJECTIVE:To screen the potential diagnostic marker genes of fibromyalgia syndrome and analyze the possible immune infiltration characteristics based on bioinformatics methods,such as weighted gene co-expression network analysis(WGCNA),and machine learning. METHODS:Gene expression profiles in peripheral serum of fibromyalgia syndrome patients and healthy controls were obtained from the gene expression omnibus(GEO)database.The differentially co-expressed genes were screened in the expression profile by differential analysis and WGCNA analysis.Least absolute shrinkage and selection operator(LASSO)and support vector machine-recursive feature elimination(SVM-RFE)machine learning algorithm were further used to identify hub biomarkers,and draw receiver operating characteristic curve(ROC)to evaluate the accuracy of diagnosing fibromyalgia syndrome.Finally,single sample gene set enrichment analysis(ssGSEA)and gene set enrichment analysis(GSEA)were used to evaluate the immune cell infiltration and pathway enrichment in patients with fibromyalgia syndrome. RESULTS AND CONCLUSION:Eight down-regulated differentially expressed genes(DEGs)were obtained after differential analysis of the GSE67311 dataset according to the conditions of log2|(FC)|>0 and P<0.05.After WGCNA analysis,497 genes were included in the module(MEdarkviolet)with the highest positive correlation(r=0.22,P=0.04),and 19 genes were included in the module(MEsalmon2)with the highest negative correlation(r=-0.41,P=6×10-5).After intersecting DEGs and the module genes of WGCNA,seven genes were obtained.Four genes were screened out by LASSO regression algorithm and five genes were screened out by SVM-RFE machine learning algorithm.After the intersection of the two,three core genes were identified,which were germinal center associated signaling and motility like,integrin beta-8,and carboxypeptidase A3.The areas under the ROC curve of the three core genes were 0.744,0.739,and 0.734,respectively,indicating that they have good diagnostic value and can be used as biomarkers for fibromyalgia syndrome.The results of immune infiltration analysis showed that memory B cells,CD56 bright NK cells,and mast cells were significantly down-regulated in patients with fibromyalgia syndrome compared with the control group(P<0.05),and were significantly positively correlated with the above three biomarkers(P<0.05).The enrichment analysis suggested that there were nine fibromyalgia syndrome enrichment pathways,mainly related to olfactory transduction pathway,neuroactive ligand-receptor interaction,and infection pathway.The above results showed that the occurrence and development of fibromyalgia syndrome are related to the involvement of multiple genes,abnormal immune regulation,and multiple pathways imbalance.However,the interactions between these genes and immune cells,as well as their relationships with various pathways need to be further investigated.

Objective To investigate the regulatory pathway of lipid peroxidation in ferroptosis and the pathogenesis of Parkinson's disease(PD)at animal level.Methods Zebrafish PD model was established by exposure to 200 μmol/L 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).Zebrafish embryo in 48 h after fertilisation were grouped into a control group(n=100),a MPTP group(n=100),and a MPTP+Nomi group(1.5 μg/ml nomifensine,n=100).Behavioral analysis was performed on zebrafish in 5 d after fertilization for movement ability,rate and distance.The expression of α-synuclein and tyrosine hydroxylase in the zebrafish brain was detected by immu-nofluorescence assay to evaluate the activity of dopaminergic neurons(DA).The levels of iron,glutathione(GSH)and lipid peroxidation,which includes catalase(CAT),glutathione peroxidase 4(GPX4),lipid peroxide(LPO),malondialdehyde(MDA)and reactive oxygen species(ROS),were measured to verify whether iron deposition may cause damage to DA through the metabolic regulation mechanism of lipid peroxidation in iron death.Results Compared with the control group,the movement rate and distance were significantly reduced in the MPTP group(P＜0.01),while Nomi treatment obviously increased the movement rate and distance when compared with the MPTP group(P＜0.01).The MPTP group exhibited obvious DA damage in comparison with the control group,and Nomi treatment reversed the damage caused by MPTP.There was no sta-tistical difference in the CAT level in the three group(P＞0.05).The MPTP group had signifi-cantly increased levels of iron,LPO and MDA and decreased levels of GSH and GPX4 when com-pared with the control group(838.18±143.42 vs 478.53±112.58,P＜0.05;1.71±0.11 vs 1.48±0.14,P＜0.05;4.50±0.64 vs 4.23±0.13,P＜0.05;38.93±1.72 vs 45.97±2.32,P＜0.05;0.17±0.03 vs 0.39±0.04,P＜0.05).The MPTP+Nomi group had significantly increased levels of GSH and GPX4 and decreased levels of iron and LPO when compared with the MPTP group(40.79±1.02 vs 38.93±1.72,P＜0.05;0.26±0.05 vs 0.17±0.03,P＜0.05;326.75±110.95 vs 838.18±143.42,P＜0.05;1.01±0.27 vs 1.71±0.11,P＜0.05).The results of the ROS flow cytometry analysis showed that the percentages of ROS positive cells in the control group,MPTP group,and Nomi group were as follows:Histogram:31.6％,46.2％and 31.3％,Scatter plot:62.4％,73.0％and 65.7％,respectively.Conclusion Ferroptosis is found in PD zebrafish induced by MPTP exposure.Our study has clarified at animal level that the damage of DA caused by iron deposition is related to the metabolic mechanism of lipid peroxidation caused by iron death.

Objective To investigate the regulatory pathway of lipid peroxidation in ferroptosis and the pathogenesis of Parkinson's disease(PD)at animal level.Methods Zebrafish PD model was established by exposure to 200 μmol/L 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).Zebrafish embryo in 48 h after fertilisation were grouped into a control group(n=100),a MPTP group(n=100),and a MPTP+Nomi group(1.5 μg/ml nomifensine,n=100).Behavioral analysis was performed on zebrafish in 5 d after fertilization for movement ability,rate and distance.The expression of α-synuclein and tyrosine hydroxylase in the zebrafish brain was detected by immu-nofluorescence assay to evaluate the activity of dopaminergic neurons(DA).The levels of iron,glutathione(GSH)and lipid peroxidation,which includes catalase(CAT),glutathione peroxidase 4(GPX4),lipid peroxide(LPO),malondialdehyde(MDA)and reactive oxygen species(ROS),were measured to verify whether iron deposition may cause damage to DA through the metabolic regulation mechanism of lipid peroxidation in iron death.Results Compared with the control group,the movement rate and distance were significantly reduced in the MPTP group(P＜0.01),while Nomi treatment obviously increased the movement rate and distance when compared with the MPTP group(P＜0.01).The MPTP group exhibited obvious DA damage in comparison with the control group,and Nomi treatment reversed the damage caused by MPTP.There was no sta-tistical difference in the CAT level in the three group(P＞0.05).The MPTP group had signifi-cantly increased levels of iron,LPO and MDA and decreased levels of GSH and GPX4 when com-pared with the control group(838.18±143.42 vs 478.53±112.58,P＜0.05;1.71±0.11 vs 1.48±0.14,P＜0.05;4.50±0.64 vs 4.23±0.13,P＜0.05;38.93±1.72 vs 45.97±2.32,P＜0.05;0.17±0.03 vs 0.39±0.04,P＜0.05).The MPTP+Nomi group had significantly increased levels of GSH and GPX4 and decreased levels of iron and LPO when compared with the MPTP group(40.79±1.02 vs 38.93±1.72,P＜0.05;0.26±0.05 vs 0.17±0.03,P＜0.05;326.75±110.95 vs 838.18±143.42,P＜0.05;1.01±0.27 vs 1.71±0.11,P＜0.05).The results of the ROS flow cytometry analysis showed that the percentages of ROS positive cells in the control group,MPTP group,and Nomi group were as follows:Histogram:31.6％,46.2％and 31.3％,Scatter plot:62.4％,73.0％and 65.7％,respectively.Conclusion Ferroptosis is found in PD zebrafish induced by MPTP exposure.Our study has clarified at animal level that the damage of DA caused by iron deposition is related to the metabolic mechanism of lipid peroxidation caused by iron death.

Objective:To investigate the situation of mild cognitive impairment (MCI) in elderly people in fluorosis areas, and analyze its potential influencing factors.Methods:From April to October 2020, a cluster random sampling method was used to select local elderly people aged 60 and older in areas of drinking-water-borne endemic fluorosis in Tianjin, and their general information was collected through face-to-face surveys. Terminal drinking water samples were collected from survey subjects. Water fluoride concentration was measured using ion-selective electrode method. Cognitive function was evaluated using the Mini-Mental State Examination (MMSE), and survey subjects were categorized into MCI group and control group based on MMSE scores. Logistic regression and Chi-squared automatic interaction detection classification tree models were constructed to analyze the influencing factors of MCI.Results:A total of 677 subjects were included, including 160 cases in the MCI group and 517 cases in the control group, with a MCI detection rate of 23.63%. The detection rates of MCI in the high fluoride exposure group and non high fluoride exposure group were 30.82% (94/305) and 17.74% (66/372), respectively, with a statistically significant difference between the groups (χ 2 = 15.88, P < 0.001). The results of univariate analysis showed that MCI was associated with high fluoride exposure, age, per capita annual household income, sleep duration, personality, and interpersonal relationships (χ 2 = 15.88, 16.17, 12.32, 8.91, 14.03, 9.81, P < 0.05). The results of multiple logistic regression analysis showed that moderate interpersonal relationships ( OR = 2.13, 95% CI: 1.08, 4.20) and high fluoride exposure ( OR = 1.91, 95% CI: 1.03, 3.54) were both risk factors for MCI ( P < 0.05). The classification tree model analysis result showed that high fluoride exposure was the primary factor affecting MCI in elderly people in fluorosis areas, followed by per capita annual household income, personality, and age. Conclusions:The detection rate of MCI in the high fluoride exposure group of elderly people in fluorosis areas is higher than that in the non high fluoride exposure group. High fluoride exposure, interpersonal relationships, per capita annual household income, personality, and age are all influencing factors of MCI in elderly people in fluorosis areas.

Objective:To learn about the iodine nutrition status of pregnant women in Tianjin and analyze the influencing factors of moderate to severe iodine deficiency in pregnant women.Methods:From January 2022 to December 2024, a stratified random sampling method was adopted. Each year, five sampling areas were divided into the east, west, south, north and central directions in 16 districts of Tianjin. Ten pregnant women from one township/street in each area were selected for questionnaire surveys. Household salt samples and random urine samples were collected to measure salt iodine and urinary iodine levels. Multivariate logistic regression was used to analyze the influencing factors of moderate to severe iodine deficiency in pregnant women, and a restricted cubic spline model was used to analyze the dose-response relationship between the risk of moderate to severe iodine deficiency in pregnant women and salt iodine level.Results:A total of 2 532 pregnant women in Tianjin were surveyed, with an age of (30.70 ± 4.44) years. Among them, 53.20% (1 347/2 532) had received health education, and 52.45% (1 328/2 532) actively supplemented iodine. A total of 2 532 household salt samples of pregnant women were tested, with a median salt iodine level of 23.66 mg/kg. The coverage rate of iodized salt was 74.45% (1 885/2 532), the qualified rate of iodized salt was 87.59% (1 651/1 885), and the consumption rate of qualified iodized salt was 65.21% (1 651/2 532). A total of 2 532 urine samples of pregnant women were tested, with a median urinary iodine level of 151.68 μg/L. Among them, 652, 1 348 and 532 urine samples were collected from pregnant women in the early, middle, and late stages of pregnancy, respectively, with median urinary iodine levels of 150.80, 153.00 and 143.68 μg/L, respectively. Multivariate logistic regression analysis showed that consuming iodized salt was a protective factor for the moderate to severe iodine deficiency in pregnant women ( OR = 0.70, 95% CI: 0.58 - 0.86, P < 0.001). Restricted cubic spline analysis showed that there was a linear dose-response relationship between the risk of moderate to severe iodine deficiency in pregnant women and salt iodine level ( Poverall < 0.001, Pnonlinear = 0.065). Further stratified analysis revealed that there was a linear dose-response relationship between the risk of moderate to severe iodine deficiency in pregnant women who had received iodine supplementation and salt iodine level ( Poverall = 0.018, Pnonlinear = 0.554), while there was a nonlinear dose-response relationship between the risk of moderate to severe iodine deficiency in pregnant women who had not received iodine supplementation and salt iodine level ( Poverall < 0.001, Pnonlinear = 0.029). Conclusions:From 2022 to 2024, the overall iodine level of pregnant women in Tianjin is appropriate, but some pregnant women are at risk of iodine deficiency. Consuming iodized salt is a protective factor against the moderate to severe iodine deficiency in pregnant women. It is still necessary to further implement comprehensive prevention and control measures mainly based on iodized salt.