Background Lead smelting workers are exposed to mixed heavy metals such as lead (Pb) and cadmium (Cd). However, the specific associations and molecular mechanisms by which their combined exposure induces genetic damage remain unclear. Objective To clarify the association between combined Pb-Cd exposure and genetic damage and to explore the possible biological mechanisms through occupational epidemiological investigations and animal experiments. Methods (1) Population study: A cross-sectional study was conducted on 374 lead smelting workers in northern China. Inductively coupled plasma mass spectrometry (ICP-MS) was used to detect urinary levels of 8 metals including Pb and Cd, and graphite furnace atomic absorption spectroscopy (GFAAS) was used to quantify blood levels of Pb and Cd. The cytokinesis-block micronucleus assay (CBMN) was used to assess genetic damage. Poisson regression was used to analyze the association between metal exposure and micronucleus rates. (2) In vivo experiment: Thirty SD rats were randomly assigned to five groups: control (pure water), Pb (300 mg·L⁻¹ lead acetate), Cd (50 mg·L⁻¹ cadmium chloride), combined exposure (Pb + Cd), and resveratrol intervention (Pb + Cd + 50 mg·L⁻¹ resveratrol). After 8 weeks of ad libitum drinking water exposure, liver pathology, oxidative stress indicators [reactive oxygen species (ROS), reduced glutathione (GSH), oxidized glutathione (GSSG), malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD)], genetic damage (Comet assay and γ-H2AX) were evaluated. Furthermore, cell cycle distribution, apoptosis rates, and mRNA expression of DNA damage response (DDR), DNA repair, and apoptosis-related genes were measured. Results (1) The geometric mean (GM, 95%CI) of urinary Pb and Cd were 14.69 (13.14, 16.51) µg·L⁻¹ and 2.11 (1.90, 2.33) µg·L⁻¹, respectively; the blood Pb and Cd levels were 117.10 (105.59, 129.87) µg·L⁻¹ and 4.55 (4.23, 4.89) µg·L⁻¹, respectively among the 374 workers. The mean micronucleus rate was (1.64±0.081) ‰, with significantly higher rates in males (1.65±0.083) ‰ than females (1.53±0.334) ‰ (U=4.166, P=0.041). All Pb and Cd biomarkers were positively correlated with micronucleus rate (FR>1, P<0.05), with a significant interaction effect observed between Pb and Cd (FR>1, P<0.05). (2) In rats, co-exposure to Pb and Cd caused liver tissue damage and inflammatory infiltration. Significant increases were observed in lymphocyte ROS; GSSG and MDA in lung tissue increased, while GSH and CAT activity decreased. Comet assay indicators and γ-H2AX levels were significantly elevated. Co-exposure induced S-phase arrest and increased apoptosis. mRNA levels of DDR (ATM, ATR, Chk2, and P53) and pro-apoptotic genes (Bax and Caspase-3) were upregulated, while the anti-apoptotic gene Bcl-2 and DNA repair genes (BRCA1, BRCA2, RAD51, RAD52, and CtIP) were downregulated. Two-way ANOVA confirmed synergistic effects on GSSG, Comet assay indicators, and ATR/Chk2 mRNA expression. Conclusion Occupational co-exposure to Pb and Cd synergistically induces genetic damage. This damage is mediated by oxidative stress and DNA damage, which activates the DDR pathway and inhibits the expression of DNA repair genes, ultimately leading to cell cycle arrest and apoptosis.

Curcuma zedoaria (commonly known as "E Zhu" in traditional Chinese medicine) is a perennial herb, and its rhizome is used for medical purpose. It is clinically effective for treating various symptoms caused by blood stasis. For oral administration, E Zhu is often combinded with other traditional Chinese medicines to prepare various formulations such as decoctions, pills, and tablets. This allows its medicinal properties to reach the internal organs and meridians, promotes the flow of qi and blood, and dissipates stasis and stagnation. E Zhu is mainly distributed in Guangxi, Sichuan, Zhejiang, Yunnan, and other regions in China. However, cadmium (Cd) contamination has emerged in multiple areas due to the development of battery, smelting, electroplating, and chemical industries. Additionally, the intrinsic Cd-accumulation property of E Zhu coupled with the Cd pollution has rendered a prominent issue during its cultivation and medicinal application, thereby comprimising the safety and efficacy of its clinical use. This paper comprehensively reviewed the absorption and accumulation patterns of Cd in E Zhu and proposed a comprehensive chain of control measures from cultivation to processing and purification, aiming to reduce Cd in E Zhu. These measures are expected to lower the potential risk of low-dose Cd accumulation in the human body and further ensure the safe clinical application of E Zhu as a raw material in traditional Chinese medicine.

Background Lead smelting workers are exposed to mixed heavy metals such as lead (Pb) and cadmium (Cd). However, the specific associations and molecular mechanisms by which their combined exposure induces genetic damage remain unclear. Objective To clarify the association between combined Pb-Cd exposure and genetic damage and to explore the possible biological mechanisms through occupational epidemiological investigations and animal experiments. Methods (1) Population study: A cross-sectional study was conducted on 374 lead smelting workers in northern China. Inductively coupled plasma mass spectrometry (ICP-MS) was used to detect urinary levels of 8 metals including Pb and Cd, and graphite furnace atomic absorption spectroscopy (GFAAS) was used to quantify blood levels of Pb and Cd. The cytokinesis-block micronucleus assay (CBMN) was used to assess genetic damage. Poisson regression was used to analyze the association between metal exposure and micronucleus rates. (2) In vivo experiment: Thirty SD rats were randomly assigned to five groups: control (pure water), Pb (300 mg·L⁻¹ lead acetate), Cd (50 mg·L⁻¹ cadmium chloride), combined exposure (Pb + Cd), and resveratrol intervention (Pb + Cd + 50 mg·L⁻¹ resveratrol). After 8 weeks of ad libitum drinking water exposure, liver pathology, oxidative stress indicators [reactive oxygen species (ROS), reduced glutathione (GSH), oxidized glutathione (GSSG), malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD)], genetic damage (Comet assay and γ-H2AX) were evaluated. Furthermore, cell cycle distribution, apoptosis rates, and mRNA expression of DNA damage response (DDR), DNA repair, and apoptosis-related genes were measured. Results (1) The geometric mean (GM, 95%CI) of urinary Pb and Cd were 14.69 (13.14, 16.51) µg·L⁻¹ and 2.11 (1.90, 2.33) µg·L⁻¹, respectively; the blood Pb and Cd levels were 117.10 (105.59, 129.87) µg·L⁻¹ and 4.55 (4.23, 4.89) µg·L⁻¹, respectively among the 374 workers. The mean micronucleus rate was (1.64±0.081) ‰, with significantly higher rates in males (1.65±0.083) ‰ than females (1.53±0.334) ‰ (U=4.166, P=0.041). All Pb and Cd biomarkers were positively correlated with micronucleus rate (FR>1, P<0.05), with a significant interaction effect observed between Pb and Cd (FR>1, P<0.05). (2) In rats, co-exposure to Pb and Cd caused liver tissue damage and inflammatory infiltration. Significant increases were observed in lymphocyte ROS; GSSG and MDA in lung tissue increased, while GSH and CAT activity decreased. Comet assay indicators and γ-H2AX levels were significantly elevated. Co-exposure induced S-phase arrest and increased apoptosis. mRNA levels of DDR (ATM, ATR, Chk2, and P53) and pro-apoptotic genes (Bax and Caspase-3) were upregulated, while the anti-apoptotic gene Bcl-2 and DNA repair genes (BRCA1, BRCA2, RAD51, RAD52, and CtIP) were downregulated. Two-way ANOVA confirmed synergistic effects on GSSG, Comet assay indicators, and ATR/Chk2 mRNA expression. Conclusion Occupational co-exposure to Pb and Cd synergistically induces genetic damage. This damage is mediated by oxidative stress and DNA damage, which activates the DDR pathway and inhibits the expression of DNA repair genes, ultimately leading to cell cycle arrest and apoptosis.

ObjectiveTo investigate the preventive and therapeutic effects of Qingfei Paidu decoction (QFPDD) on acute lung injury (ALI) in mice and its underlying molecular mechanisms based on miRNA sequencing technology. MethodsTwenty-four 4-week-old male KM mice were randomly divided into a control group, a model group, and a QFPDD group (n = 8 per group). After one week of acclimatization, mice in the control and model groups were intragastrically administered ultrapure water (0.2 mL per dose), whereas mice in the QFPDD group were intragastrically administered QFPDD (1.6 g crude drug/mL, 0.2 mL per dose), twice daily for 8 consecutive days. On days 2–8, mice in the model and QFPDD groups were exposed to aerosolized lipopolysaccharide (LPS) solution (2.5 g/L, 4 mL per exposure) for 7 consecutive days. On day 9, blood was collected via the retro-orbital venous plexus under deep anesthesia, and lung tissues were harvested. Body weight and lung weight were measured, and the lung coefficient was calculated. Serum levels of inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 were detected by ELISA. Lung histopathological changes were observed by HE staining of paraffin-embedded sections. miRNA expression profiles in lung tissues were analyzed using the Illumina HiSeq 2500 sequencing platform. Target genes of differentially expressed miRNAs were predicted using bioinformatics databases, and functional enrichment analysis of these target genes was performed using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Differentially expressed miRNAs were validated by reverse transcription quantitative real-time PCR (RT-qPCR). ResultsCompared with the control group, the model group showed a consistent body weight growth trend but a significantly increased lung coefficient (P < 0.01). ELISA results showed that serum levels of TNF-α and IL-6 were significantly elevated in the model group compared with the control group (P < 0.01), whereas QFPDD treatment significantly reduced serum TNF-α and IL-6 levels compared with the model group (P < 0.05). HE staining showed that, compared with the control group, the model group exhibited widened alveolar septa, massive inflammatory cell infiltration, partial alveolar expansion, and mild capillary dilation with congestion. In contrast, the QFPDD group showed only slightly widened alveolar septa and mild inflammatory cell infiltration compared with the model group. Intersection analysis of miRNA sequencing data identified 13 differentially expressed miRNAs common to both the model vs. control and QFPDD vs. model comparisons. Among them, 6 miRNAs (mmu-miR-203-3p, mmu-miR-181b-5p_R-1, hsa-miR-4286_R+1, mmu-miR-1843b-5p_L+1R-1_2, mmu-miR-22-3p, and mmu-miR-1964-3p) were significantly up-regulated in the model group (P < 0.05) and significantly down-regulated after QFPDD treatment (P < 0.05), showing a therapeutic reversal trend. GO analysis revealed that the target genes of the differentially expressed miRNAs were mainly enriched in biological processes such as RNA polymerase Ⅱ transcriptional regulation. KEGG analysis indicated that target genes were mainly enriched in signaling pathways including the mitogen-activated protein kinase (MAPK) pathway. RT-qPCR validation result for mmu-miR-203-3p was consistent with the sequencing analysis results. ConclusionQFPDD may exert preventive and therapeutic effects against ALI by regulating the expression of mmu-miR-203-3p and other miRNAs, thereby modulating inflammatory responses and the MAPK signaling pathway and participating in the pathological process of lung injury.

Objective:To explore the clinical efficacy of targeted epidural space block combined with acupotomy in the treatment for lumbar disc herniation(LDH)under the monitor of X-ray image,and to analyze the safety of its operation.Methods:A total of 60 LDH patients who admitted to Department of Pain,Jiangyin Affiliated Hospital of Nanjing University of Chinese Medicine from January 2022 to September 2023 were selected,and they were randomly divided into observation group and control group as random number table,with 30 cases in each group.The observation group adopted targeted epidural space block combined with acupotomy under monitor with X-ray for anterior epidural space,while the control group adopted conventionally epidural block,all of which were treated only once.The Visual Analogue Scale(VAS)scores and Oswestry Disability Index(ODI)scores of two groups before treatment and at the 3rd day,7th day and 30th day after treatment were compared.At the 30th day after treatment,the curative effects of two groups were evaluated according to ODI scores,at the same time,the safe distance between puncture and acupotomy in patients of observation group was analyzed.Results:The effective rates of the treatment of observation group and control group were respectively 96.67%and 70%,and the difference of that between the two groups was statistically significant(x2=7.680,P<0.05).The VAS scores of the two groups at the 3rd day,7th day and 30th day after treatment were lower than those before treatment,and the observation group(2.30±0.87),(1.53±0.86)and(1.47±0.81)were lower than the control group(4.43±0.89),(2.23±0.89)and(4.27±0.82),the difference was statistically significant(t=9.374,3.098,13.306,P<0.05).The ODI scores of the two groups were at the 3rd day,7th day and 30th day after treatment were lower than those before treatment,and the differences of them between two groups were statistically significant(t=7.112,10.690,10.151,P<0.05).In 30 patients of observation group,the gap between the dural sac and the inner edge of facet joint on the affected side was larger than 4.0mm in 25 cases,and all of them can safely conduct puncture with targeting.Conclusion:The targeted epidural space block combined with acupotomy by adopting monitor with X-ray image can effectively improve the symptom of pain,and activities of daily life of LDH patients,which treatment is safe and effective.

Objective To explore the diagnostic value of brain cord blood flow ultrasound combined with glucose metabolism indexes in the patients with preeclampsia(PE)complicated with fetal growth restriction(FGR).Methods This study was a prospective cohort case-control study.A total of 198 PE patients admitted to the hospital from January 2017 to January 2022 were collected and divided into group A(PE patients com-plicated with FGR,101 cases)and group B(PE patients without FGR,97 cases)according to whether they were complicated with FGR.45 pregnant women with healthy prenatal examination in the hospital were se-lected as group C,and the differences of brain cord blood flow ultrasound indexes and glucose metabolism inde-xes among the three groups and the efficiency for diagnosing PE complicated with FGR were analyzed.Results Fast-ing blood glucose and fasting insulin levels in group A,group B and group C were gradually decreased(P＜0.05).The peak systolic flow velocity/peak diastolic flow velocity(S/D),resistance index(RI)and pulsatility index(PI)of middle cerebral artery(MCA)of fetus in group A,group B and group C increased gradually(P＜0.05).The levels of umbilical artery(UA)of fetus in group A,group B and group C decreased gradually(P＜0.05).The area under the curve of fasting insulin,fasting blood glucose,S/D,RI and PI of MCA,S/D,RI,PI of UA and the combined detection were 0.782,0.748,0.765,0.836,0.805,0.758,0.811,0.795 and 0.895,respectively.The diagnostic efficiency of combined detection was higher than that of single diagnosis(Z=4.327,4.862,4.551,3.400,3.946,4.679,3.742,4.102,P＜0.05).Conclusion The combined detection of glucose metabolism indexes and brain cord blood flow ultrasound indexes is of great value in the differential diagnosis of patients with PE complicated with FGR,and has good clinical application value.

Objective To assess stromal fibrosis in epidermal growth factor receptor(EGFR)mutant lung adenocarcinoma and its association with resistance to targeted therapy and patient prognosis.Methods Medical records of 207 patients diagnosed with EGFR-mutant advanced lung adenocarcinoma who received treatment at a hospital between January 2021 and December 2022 were reviewed.A total of 86 patients were ultimately included based on their prognosis and survival duration.These patients were categorized into a resistance group(32 cases)and a non-resistance group(54 cases),depending on whether they developed resistance to targeted therapy within one year.Additionally,patients were classified into mild,moderate,and severe fibrosis groups according to the extent of fibrosis observed.Clinical and pathological characteristics,as well as fibrosis levels,were compared between the two groups.Factors influencing the development of resistance to targeted therapy in patients with EGFR-mutant lung adenocarcinoma were analyzed,and the survival outcomes of patients with varying degrees of fibrosis were evaluated during follow-up.Results In the resistance group,the prevalence of EGFR exon 20 insertion mutations,elevated CA125 levels,and the presence of moderate-to-severe fibrosis were significantly higher compared to the non-resistance group(P<0.05).Multivariate logistic regression analysis revealed that EGFR exon 20 inser-tion mutation(OR=3.691,95%CI:1.043～13.057),elevated CA125 levels(OR=4.104,95%CI:1.160～14.517),and moderate-to-severe fibrosis(OR=3.959,95%CI:1.410～11.115)were independent risk factors associated with resistance to targeted therapy among patients with EGFR-mutant lung adenocarcinoma(P<0.05).The Cox proportional hazards model demonstrated a C-index of 0.72(95%CI:0.65～0.79),with area under the curve(AUC)values for 1-year and 2-year survival predictions of 0.781 and 0.734,respectively.EGFR exon 20 insertion mutation(HR=3.691),moderate-to-severe fibrosis(HR=3.959),and elevated CA125 levels(HR=4.104)were identified as independent prognostic factors for overall survival in these patients following targeted therapy.The median progression-free survival(PFS)for patients with mild,moderate,and severe fibrosis was 10.5 months,7.2 months,and 3.9 months,respectively,while the median overall survival(OS)was 21.4 months,16.1 months,and 11.5 months,respectively.Statistically significant differences in both PFS and OS were observed across the three fibrosis severity groups.(P<0.05).Conclusion The extent of stromal fibrosis in EGFR-mutant lung adenocarcinoma influences resistance to targeted therapy,and the progression of fibrosis is correlated with an unfavorable prognosis.

Objective:To explore the role of the expression of peroxisome proliferator-activated receptor β/δ(PPARβ/δ)in the anti-inflammatory mechanism of acupuncture(AC)against cerebral ischemia-reperfusion injury(CIRI)in rat hippocampal tissue.Methods:A total of 60 SD rats were randomly divided into Sham group,CIRI group,CIRI+AC group and CIRI+AC+PPARβ/δsiRNA group.The CIRI rat model was prepared by the method of middle cerebral artery occlusion,and acupuncture treatment was used for intervention.The neurological function of rats was assessed by the modified neurological severity score(mNSS)method after 72 hours of intervention.The brain in-farction volume ratio was observed by 2,3,5-triphenyltetrazolium chloride(TTC)staining method.The pathological morphological changes of brain tissue were observed by hematoxylin-eosin(HE)staining.The expression levels of PPARβ/δ,IL-1β,and IL-10 mRNA in the hippocampal region were detected by RT-qPCR.Results:Compared with the Sham group,the mNSS scores and the brain infarction volume ratio in all modeling groups were significantly in-creased.Compared with the CIRI group,the mNSS scores and the brain infarction volume ratio in the CIRI+AC group were significantly reduced.The expression levels of PPARβ/δ and IL-10 in the hippocampus were significantly upregu-lated,while the expression level of IL-1β was significantly downregulated(P<0.01),with scattered inflammatory cell infiltration observed.Compared with the CIRI+AC group,there was no significant difference in the mNSS scores and the brain infarction volume ratio in the CIRI+AC+PPARβ/δsiRNA group.The expression level of PPARβ/δ in the hippocampus was significantly downregulated(P<0.01),while no significant difference was found in the expression level of IL-10 and IL-1β mRNA,with a small amount of inflammatory cell infiltration observed.Conclusion:Acupunc-ture ameliorates the recovery of neurological function and the ratio of cerebral infarction volume in rats with CIRI,and its mechanism may be associated with the upregulation of PPARβ/δ expression,thereby reducing inflammatory respon-ses and exerting neuroprotective effects.

Objective:To investigate the effects of acupuncture intervention on the expression of circRNA_017109 and miR-138-5p in the hippocampal region of rats with cerebral ischemia-reperfusion injury(CIRI),and to explore the potential mechanism of acupuncture in treating CIRI.Methods:Forty-five Sprague-Dawley(SD)rats were divided into sham group,middle cerebral artery occlusion/reperfusion(MCAO/R)group,and acupuncture(MCAO/R+AC)group.The MCAO/R and MCAO/R+AC groups were subjected to the modified Longa suture method to establish the MCAO/R model.The MCAO/R+AC group received acupuncture at the"Dazhui","Renzhong",and"Baihui"acu-points.Neurological impairment was assessed via the modified neurological severity score(mNSS).while cerebral infarct volume was assessed through TTC staining.Histological alterations in brain tissue were examined via HE stai-ning,and the expression of circRNA_017109 and miR-138-5p in the hippocampal area were determined using RT-qPCR.Results:The mNSS scores and cerebral infarct volume ratios in the model groups were markedly higher than those in the sham group(P<0.01),but these were significantly reduced in MCAO/R+AC group compared to the MCAO/R group(P<0.01).HE staining showed severe necrosis and inflammation in the MCAO/R group's hippocam-pus,while the MCAO/R+AC group had minimal necrosis and scattered inflammation.RT-qPCR results indicated that the expression of circRNA_017109 and miR-138-5p was downregulated in the MCAO/R group,while MCAO/R+AC group upregulated the expression of circRNA_017109 and miR-138-5p(P<0.05,P<0.01).Conclusion:Acupunc-ture can improve neurological deficits,reduce cerebral infarction volume,and alleviate inflammatory cell infiltration in CIRI rats by upregulating the expression of circRNA_017109 and miR-138-5p,thereby exerting neuroprotective effects.

Objective: To explore glymphatic impairment in pediatric refractory epilepsy (RE) using multi-parameter magnetic resonance imaging (MRI), assess its relationship with white-matter (WM) abnormalities and clinical indicators, and preliminarily evaluate the performance of multi-parameter MRI in discriminating RE from drug-sensitive epilepsy (DSE). Materials and Methods: We retrospectively included 70 patients with DSE (mean age, 9.7 ± 3.5 years; male:female, 37:33) and 26 patients with RE (9.0 ± 2.9 years; male:female, 12:14). The diffusion tensor imaging analysis along the perivascular space (DTI-ALPS) index as well as fractional anisotropy (FA), mean diffusivity (MD), and nodal efficiency values were measured and compared between patients with RE and DSE. With sex and age as covariables, differences in the FA and MD values were analyzed using tract-based spatial statistics, and nodal efficiency was analyzed using a linear model. Pearson’s partial correlation was analyzed. Receiver operating characteristic (ROC) curves were used to evaluate the discrimination performance of the MRI-based machine-learning models through five-fold cross-validation. Results: In the RE group, FA decreased and MD increased in comparison with the corresponding values in the DSE group, and these differences mainly involved the callosum, right and left corona radiata, inferior and superior longitudinal fasciculus, and posterior thalamic radiation (threshold-free cluster enhancement, P < 0.05). The RE group also showed reduced nodal efficiency, which mainly involved the limbic system, default mode network, and visual network (false discovery rate, P < 0.05), and significantly lower DTI-ALPS index (F = 2.0, P = 0.049). The DTI-ALPS index was positively correlated with FA (0.25 ≤ r ≤ 0.32) and nodal efficiency (0.22 ≤ r ≤ 0.37), and was negatively correlated with the MD (-0.24 ≤ r≤ -0.34) and seizure frequency (r = -0.47). A machine-learning model combining DTI-ALPS, FA, MD, and nodal efficiency achieved a cross-validated ROC curve area of 0.83 (sensitivity, 78.2%; specificity, 84.8%). Conclusion Pediatric patients with RE showed impaired glymphatic function in comparison with patients with DSE, which was correlated with WM abnormalities and seizure frequency. Multi-parameter MRI may be feasible for distinguishing RE from DSE.