Trichloroethylene (TCE) is a common industrial organic solvent. Exposure to TCE can induce occupational medicamentosa-like dermatitis (OMDT), which is frequently accompanied by liver and kidney injury. During the organ damage caused by TCE, mtDNA is a key molecule that mediates the interaction between the liver and the kidney. Our previous research has shown that mitochondrial DNA (mtDNA) released from hepatocytes damage under TCE sensitization can cause damage to renal tubular epithelial cells. However, the specific mechanism remains unclear. To this end, we established a TCE percutaneous sensitization mouse model. We found that in the TCE sensitization positive group, liver damage was strongly correlated with kidney damage. The Stimulator of Interferon Genes (STING) pathway in renal macrophages was activated, and the macrophages underwent M1 polarization accompanied by the formation of macrophage extracellular traps (METs). To further confirm the role of hepatocyte mtDNA, we extracted the mtDNA from the hepatocytes of TCE sensitization positive mice and injected it into normal mice. We found that the STING pathway in the kidney of the injection group was activated, and the macrophages also underwent M1 polarization. This phenomenon was significantly alleviated after intervention with the STING inhibitor C-176. We also confirmed this in vitro using RAW264.7 with hepatocyte mtDNA treatment. In summary, our work demonstrated that hepatocyte mtDNA can activate the STING pathway in renal macrophages, thereby causing M1 polarization of macrophages and the formation of METs. C-176 can alleviate this phenomenon.

ObjectivesNeutrophil extracellular traps (NETs) are a crucial mechanism of the neutrophil immune response and exhibit complex dual pathological and physiological effects in pulmonary diseases. In recent years, this research domain has increased significantly, yet systematic bibliometric research remains absent. Therefore, this study utilizes bibliometric methods to thoroughly examine the research landscape and development sequence of this domain, aiming to provide theoretical references for future research.MethodsIn this study, VOSviewer and CiteSpace were used to conduct a visualization analysis of the article concerning NETs and pulmonary diseases indexed in the Web of Science Core Collection (WoSCC) and Scopus databases from January 1st, 2006, to December 31st, 2025. Additionally, we utilized the R package bibliometric and Origin 2025b to optimize the data and visualization charts, ensuring that the analytical results were presented with greater clarity and intuitiveness.ResultsThis study covered 1, 539 articles from 561 journals. The results indicate that since 2006, this domain has seen a phased growth trend. The phase from 2010 to 2018 was a stable growth phase; After 2019, propelled by the COVID-19 virus infection (COVID-19), it entered a significant development phase, and reached its peak in 2022. In terms of influence, China and the United States are leading contributors. Mark R. Looney of the University of California is the core author, and “Frontiers in Immunology” is the most frequently cited journal. Through the visualization analysis of topic categories, keywords, and references indicates that research focal points in this domain have progressively transitioned from initial fundamental mechanisms and pathological descriptions to concerns regarding pulmonary inflammation, immune thrombosis, COVID-19, and biomarkers, subsequently extending to frontiers such as the NLRP3 inflammasome, interstitial lung disease (ILD), and tumor microenvironment (TME).ConclusionsThis study is the first application of bibliometric methods to visually present research domains related to NETs and pulmonary diseases, revealing the correlation between NETs and ILD, lung cancer (LC), and respiratory infections, along with the underlying mechanisms, continues to be a hot topic in research. However, the predominant function of NETs in specific diseases and their potential utility as therapeutic targets still necessitate further systematic elaboration.

In this study, a novel pyrazole amide derivative designated T1, was designed and synthesized to explore its therapeutic potential for liver cancer. The compound was evaluated for its effects on liver cancer cell viability, proliferation, migration, invasion, and apoptosis both in vitro and in vivo. The in vitro evaluation was conducted using several experimental techniques, including the CCK-8 assay, colony formation assay, transwell migration assay, immunofluorescence, and western blotting. T1 demonstrated an IC50 of 47.7 μg/mL in HuH7 liver cancer cells, showing significantly improved efficacy compared to standard chemotherapeutics, including capecitabine and 5-fluorouracil, and comparable efficacy to sorafenib (IC50 of 55.6 μg/ml). Furthermore, T1 significantly inhibited the proliferation, migration, and invasion of HepG2 and HuH-7 cells, while also inducing apoptosis. Transcriptome sequencing and western blotting analyses revealed that T1 modulates the MAPK signaling pathway, leading to the altered expression of apoptosis-related proteins, which contributes to the observed anti-cancer effects. In vivo studies using a HuH-7 xenograft mouse model confirmed the efficacy of T1, with tumor growth inhibition similar to that of sorafenib. These findings suggest that T1 holds significant promise as a potential chemotherapeutic agent for the treatment of hepatocellular carcinoma (HCC).

The bacterial genome contains numerous repeated sequences that greatly affect its genomic plasticity. The Escherichia coli K-12 genome contains three copies of the TRIP1 repeat sequence (TRIP1a, TRIP1b, and TRIP1c). However, the diversity, distribution, and role of the TRIP1 repeat sequence in the E. coli genome are still unclear. In this study, after screening 6725 E. coli genomes, the TRIP1 repeat was found in the majority of E. coli strains (96%: 6454/6725). The copy number and direction of the TRIP1 repeat sequence varied in each genome. Overall, 2449 genomes (36%: 2449/6725) had three copies of TRIP1 (TRIP1a, TRIP1b, and TRIP1c), which is the same as E. coli K-12. Five types of TRIP1 repeats, including two new types (TRIP1d and TRIP1e), are identified in E. coli genomes, located in 4703, 3529, 5741, 1565, and 232 genomes, respectively. Each type of TRIP1 repeat is localized to a specific locus on the chromosome. TRIP1 repeats can cause intra-chromosomal rearrangements. A total of 156 rearrangement events were identified, of which 88% (137/156) were between TRIP1a and TRIP1c. These findings have important implications for future research on TRIP1 repeats.

AimThis study aimed to explore the value of T1 mapping in assessing the grade and stage of rectal adenocarcinoma and its correlation with tumor tissue composition.MethodsInformed consent was obtained from all rectal cancer patients after approval by the institutional review board. Twenty-four patients (14 women and 10 men; mean age, 64.46 years; range, 35 – 82 years) were enrolled in this prospective study. MRI examinations were performed using 3.0T MR scanner before surgery. HE, immunohistochemical, and masson trichrome-staining was performed on the surgically resected tumors to assess the degree of differentiation, stage, and invasion. Two radiologists independently analyzed native T1 and postcontrast T1 for each lesion, and calculated the extracellular volume (ECV) was calculated from T1 values. Intraclass correlation coefficient (ICC) and Bland-Altman plots were applied to analyze the interobserver agreement of native T1 values and postcontrast T1 values. Student’s t-test and one-way analysis of variance (ANOVA) were used to test the differences between T1 mapping parameters and differentiation types, T and N stages, and venous and neural invasion. Pearson correlation coefficients were used to analyze the correlation of T1 mapping extraction parameters with caudal type homeobox 2 (CDX-2), Ki-67 index, and collagen expression.ResultsBoth the native and postcontrast T1 values had an excellent interobserver agreement (ICC 0.945 and 0.942, respectively). Postcontrast T1 values indicated significant differences in venous invasion (t=2.497, p=0.021) and neural invasion (t=2.254, p=0.034). Pearson’s correlation analysis showed a significant positive correlation between native T1 values and Ki-67 (r=-0.407, p=0.049). There was a significant positive correlation between ECV and collagen expression (r=0.811, p=.000) and a significant negative correlation between ECV and CDX-2 (r=-0.465, p=0.022) and Ki-67 (r=-0.549, p=0.005).ConclusionPostcontrast T1 value can be used to assess venous and neural invasion in rectal cancer. ECV measurements based on T1 mapping can be used to identify cells and collagen fibers in rectal cancer.

The type VI secretion system (T6SS) and hemolysin HlyA are important virulence factors in Vibrio cholerae. The forkhead-associated (FHA) domain is a conserved phosphopeptide binding domain that exists in many regulatory modules. The FHA domain protein-encoding gene is conserved in the T6SS gene cluster and regulates the assembly and secretion of the T6SS. This study shows for the first time that the FHA domain protein TagH plays a role in controlling the hemolytic activity of V. cholerae, in addition to regulating the T6SS. TagH negatively regulates HlyA expression at the transcriptional and post-translational levels. The phosphopeptide binding sites of the FHA domain of TagH play a key role in the regulation of hemolytic activity. The deletion of tagH enhances the intestinal pathogenicity and extraintestinal invasion ability of V. cholerae, which mainly depend on the expression of HlyA. This study provides evidence that helps unravel the novel regulatory role of TagH in HlyA and provides critical insights which will aid in the development of strategies to manage HlyA.

In order to acquire both expanded binding ability with the EphA2 receptor and superior drug delivery capacity, we designed and synthesized the modified GO-SWL-Ahx-K-SWL conjugate as a potential targeted therapeutic drug for non-small cell lung cancer (NSCLC). Various characterization methods have confirmed that the conjugate is consistent with the theoretical peptide. The cytotoxicity test results showed that the conjugate was slightly more toxic to A549 cells than in 3 T3 cells, and the toxicity increased in a concentration-dependent manner. Single photon emission computed tomography/computed tomography (SPECT/CT) fusion imaging was performed to evaluate the conjugate binding to EphA2 receptor in vivo. The images showed obvious radioactive concentration in tumor tissues and significantly higher ratios of the tumor and muscle in the I-125-GO-SWL-Ahx-K-SWL group (10.78) than in the I-125-SWL-Ahx-K-SWL group (5.21) at all three time points (P < 0.01).

Spastic cerebral palsy is mainly caused by dyskinesia and postural abnormality caused by damage of pyramidal tract system. Clinically, the main manifestations are hypertonia, limb joint movement disorder, postural and gait abnormality. At present, Western medicine has not yet developed effective drugs to treat spastic cerebral palsy. In view of the problem that the incidence of spastic cerebral palsy in children is increasing year by year, this paper puts forward the treatment methods of spastic cerebral palsy in children based on the theory of "liver, spleen and kidney". The main treatment methods are to strengthen spleen, replenish qi, soften liver and calm wind or nourish liver and kidney, dredge tendons and dredge collaterals. The clinical effect of tonifying deficiency and eliminating pathogenic factors is exact. Attached case 1 is to support it.

Trichloroethylene (TCE) is a serious health hazard for workers with daily exposure, causing occupational medicamentosa-like dermatitis due to TCE (OMDT) and glomerular damage. Recent studies suggest that mTORC1 signaling is activated in various glomerular disorders; however, the role of mTORC1 signaling in TCE-induced glomerular damage remains to be explored. In the present study, 6 OMDT patients were enrolled and a TCE-sensitized mouse model was established to investigate molecular mechanisms underlying the glomerular damage associated with OMDT. Glomerular damage was assessed by levels of urine nephrin, H&E staining, and renal function test. Ultrastructural change of podocyte was investigated by transmission electron microscopy. The podocyte-related molecules including nephrin, α-actinin-4, and integrin β1 were visualized by immunofluorescence. The activation of mTORC1 signaling was confirmed by Western blot. Glomerular apoptosis was examined by the TUNEL test and Western blotting. Expression and location of cathepsin L (CTSL) were assessed by RT-PCR and immunofluorescence. Our results showed that TCE sensitization caused damage to glomerular structural integrity and also increased the activation of mTORC1 signaling, which was accompanied by podocyte loss, hypertrophy, and glomerular apoptosis. Importantly, we also found that over-expressed CTSL was mainly located in podocyte and CTSL inhibition could partially block the activation of mTORC1 signaling. Thus, our findings suggested a novel mechanism whereby hyperactive mTOR signaling contributes to TCE sensitization–induced and immune-mediated glomerular damage via CTSL activation.

Introduction This study aimed to investigate the mechanism in regulating the cross talk between glomerular endothelial cells and podocytes in “occupational medicamentosa-like dermatitis induced by trichloroethylene (OMLDT)” patients. Methods Totally 6 OMLDT patients, 18 controls, and 102 BALB/c female mice were involved in this study. Patient’s serum endothelin-1 (ET-1), angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2), blood urea nitrogen (BUN), and podocalyxin (PCX) were detected. All the mice were used to establish the trichloroethylene (TCE) sensitized mouse model. Transmission electron microscope results were used to reflect renal glomerulus injury. Protein levels were detected by Western blot. Ang-1/Ang-2 gene level was reflected by RT-PCR. Cell apoptosis level was detected by using TUNEL assay kit. Results We found that in OMLDT patients, ET-1, Ang-2, BUN, and PCX were highly expressed but Ang-1 was inhibited. In TCE sensitized positive mouse, the downregulation of Ang-1, pTie-2 and the upregulation of Ang-2 were mediated by ET-1/ETAR but not ET-1/ETBR. The promotor of apoptosis proteins was downregulated and the inhibitor of apoptosis proteins was upregulated by treating with BQ123. Discussion ET-1/ETAR-Angs/Tie-2 pathway mediated the cross talk between glomerular endothelial cells and podocytes. BQ123 can alleviate glomerulus immune injury.

采用GB 5009.11—2014《食品安全国家标准食品中总砷及无机砷的测定》中液相色谱—电感耦合等离子体质谱法测定毛毛鱼中的无机砷含量,参照CNAS—GL006《化学分析中不确定度的评估指南》和JJF 1059.1—2012《测量不确定度的评定与表示》建立数学模型,对毛毛鱼中As(III)、As(V)的精密度、回收率进行验证,同时对试验过程中带来的不确定度分量进行了来源分析和评定.试验结果表明:当毛毛鱼中无机砷含量为0.096 mg/kg时,无机砷i-As在95％的置信区间下扩展不确定度为0.007,测量过程中的不确定度最主要来源为校准曲线拟合和样品加标回收率.

Megacystis-microcolon-intestinal-hypoperistalsis syndrome (MMIHS) is a rare and severe disorder characterized by functional obstruction in the urinary and gastrointestinal tract. The molecular basis of this condition has been defined recently. Heterozygous variants in ACTG2, homozygous mutations in LMOD1, MYLK, and MYH9 were related to the pathogenesis of the syndrome, which encodes proteins involved in the process of smooth muscle contraction, supporting a myopathic basis for the disease. Recent studies have identified homozygous or compound heterozygous variants in MYH11 as a candidate gene of MMIHS. In this report, we described a nonconsanguineous Chinese family with three male fetuses affected with megacystis. Trio-targeted exome sequencing identified compound heterozygous variants, c.2051 G > A (p.R684H) and c.3540_3541delinsTT (p.(E1180D, Q1181Ter)), in MYH11 (NM_001040114). The variants were inherited from the parents, respectively. Western blotting showed a marked decrease in MYH11 protein in the proband's umbilical cord tissue compared with the control sample. The study's results confirmed that MYH11 is a candidate gene for MMIHS with autosomal recessive (AR) inheritance and expanded the mutation spectrum for this clinical condition. Combining clinical phenotype with molecular diagnosis may enable the identification of candidate genes for potential monogenic diseases and facilitate accurate genetic counseling, informed decision-making, and prenatal diagnosis.

Background: Botulinum neurotoxins (BoNTs) are among the deadliest toxins known to humans. To date, there are no approved drugs to treat botulism after intoxication of the motor neuron. Intensive efforts have been made to develop small-molecule inhibitors, however, few possess noteworthy in vivo efficacy.Methods: The primary rat cerebellar granule neuron (CGN)-based screening was performed to discover novel effective inhibitors against BoNT/A in neural cells. The anti-botulism mechanism of embelin was preliminary studied in the impact of embelin on the main steps of BoNTs intoxication and intracellular duration. The anti-botulism efficacy of natural product embelin on botulinum toxin type A and type B was assessed in BALB/c mouse.Findings: We discovered a promising anti-BoNTs natural poduct embelin, which could protect 80% and 60% of the mice against lethal dose of BoNT/A or BoNT/B at 1 h after toxin exposure, respectively. Moreover, at 8 h after toxin exposure, embelin still protect 30% and 35% of the mice against lethal dose of BoNT/A or BoNT/B. In addition, mechanism analysis proved that embelin could significantly speed up the clearance of BoNTs in neurons.Interpretation: We discovered a promising anti-BoNTs natural product embelin, which could protect mice against lethal dose of BoNTs. The novel botulism antidote will be useful for reducing the morbidity of BoNTs intoxication and have potential biodefense implications.Funding Statement: We acknowledge this work was supported by National Key R&D Program of China (2017YFC1200905), National Key Basic Research Program (973) of China (2015CB554202), and the Beijing Nova Program (Z181100006218111) for funding.Declaration of Interests: The authors declare no competing financial interest.Ethics Approval Statement: The Beijing Institute of Microbiology and Epidemiology Animal Care and Use Committee approved our study protocol. The animal care and use protocol adhered to the regulations of the Institutional Animal Care and Use Committee (IACUC).

We have previously shown trichloroethylene (TCE) induced occupational medicamentosa-like dermatitis due to TCE (OMLDT) with immune liver injury, and kallikrein-kinin system (KKS) activation as a probably mechanism underlying the immune damage. Bradykinin (BK) is an important active component of KKS system function, but the specific role of BK in the immune liver injury has never been examined. The present study aimed to explore the important role of BK and mechanisms of action in immune liver injury induced by TCE. TCE sensitization significantly increased the expression of BK receptor (B2R) in the liver. Compared to blank and vehicle control group, TCE sensitization positive mice developed exacerbated liver injury evidenced by elevated AST, ALT levels and hepatocyte damage. TCE sensitization also stimulated MAPK and STAT3 activation in liver tissue. B2R antagonist HOE140 ameliorated these changes. Kupffer cells (KCs) of the liver were also activated following TCE sensitization; both CD68+ KCs and CD16/CD32+ M1 type KCs were increased in TCE positive group. Further experiments isolated the KCs from the liver in each group and showed that TCE sensitization resulted activation of MAPK signal pathway which in turn caused release of the pro-inflammatory cytokines, IL-1β, IL-6, TNF-α, in KCs; the antagonist HOE140 again decreased these changes in KCs. These results uncover a novel role of BK and B2R cross-talk in KCs activation in TCE sensitized mice, mediated by pro-inflammatory cytokine release via MAPK and STAT3 activation, contributing to the immune liver injury.

Stenotrophomonas maltophilia (S. maltophilia), a multi-drug resistant opportunistic pathogen, is associated with nosocomial and community-acquired infections. Preventive and therapeutic strategies for such infections are greatly needed. In this study, sequence alignment analysis revealed that Outer membrane protein A (OmpA) was highly conserved among S. maltophilia strains but shared no significant similarity with human and mouse proteomes. In mice, intranasal immunization with S. maltophilia recombinant OmpA (rOmpA) without additional adjuvant induced sustained mucosal and systemic rOmpA-specific antibody responses. Treatment with rOmpA stimulated significantly higher levels of secretion of IFN-γ, IL-2, and IL-17A (All P<0.05) from the primary splenocytes isolated from rOmpA-immunized mice than from the primary splenocytes isolated from PBS-immunized mice. Furthermore, mice immunized with rOmpA showed significantly reduced bacterial burden in the lung and reduced levels of pro-inflammatory cytokines (TNF-α and IL-6) in bronchoalveolar lavage fluid (BALF) 24 hours after intranasal S. maltophilia infection, indicating that immunization with rOmpA may have protective effects against S. maltophilia challenge in mice. Our findings suggest that intranasal immunization with rOmpA may induce mucosal and systemic immune responses in mice, trigger Th1- and Th17-mediated cellular immune responses, and thus stimulate host immune defense against S. maltophilia infection. These results also demonstrate that intranasal vaccination may offer an alternative approach to current strategies since it induces a mucosal as well as a systemic immune response.

The role of environmental factors in autoimmune diseases has been increasingly recognized. While major advance has been made in understanding biological pathogen-induced autoimmune diseases, chemically triggered autoimmunity is poorly understood. Trichloroethylene (TCE), a common environmental pollutant, has recently been shown to induce autoimmunity. This study explored whether TCE could cause imbalance of T helper (Th) cell subsets which would contribute to the pathogenesis of TCE-induced medicamentosa-like dermatitis. BALB/c mice were treated with TCE via drinking water at doses of 2.5 or 5.0 mg/mL for 2, 4, 8, 12, and 16 weeks. Trichloroethylene exposure caused time- and dose-dependent increase in Th1, Th2, and Th17 and decrease in regulatory cell (Treg) in the spleen at 2, 4, 8, 12, and 16 weeks, with greatest changes mainly at 4 weeks. These effects were mirrored by similar changes in the expression of their corresponding cytokines interferon-γ, interleukin 4 (IL-4), IL-17A, and IL-10. Mechanistically, these phenotypic changes were accounted for by alterations to their respective master transcription factors T-box expressed in T cells, GATA-binding protein 3, Retinoic acid-related orphan receptor ct (RORct), and forkhead box P3. Of interest, TCE treatment shifted the ratios of Th1/Th2 and Th17/Treg; specifically, TCE increased Th17/Treg. These findings provide the first evidence that TCE exposure significantly changes the Th1/Th2/Th17/Treg paradigm and their specific cytokines driven by altered master transcription factors. This may promote autoimmune reactions in the pathogenesis of TCE-induced skin sensitization and associated damage to other tissues.

Background: Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease. Impairment of kidney functions in RA was observed. However, the mechanism of kidney injury of RA has not been clear. Plasma kallikrein-kinin system (KKS) was involved in inflammatory processes in kidney disease. Aim: This study aimed to explore the role of plasma KKS in immune reactions and kidney injury of RA. Results: The paw of AA rats appeared to be swelling and redness, the arthritis index was significantly increased on the 18, 21 and 24 d after injection and secondary inflammation in multi-sites was observed. Kidney dysfunction accompanied with inflammatory cell infiltration, tubular epithelial cell mitochondrial swelling and vacuolar degeneration, renal glomerular foot process fusions and glomerular basement membrane thickening were observed in AA rats. The expressions of neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (Kim-1) in kidney of AA rats were increased. In addition, expressions of BK, PK, B1R and B2R in the renal tissue of AA rats were up-regulated. Pro-inflammatory cytokines IL-2, IFN-gamma and TNF-alpha were increased and anti-inflammatory cytokines IL-4 and IL-10 were low in kidney. Plasma kallikrein (PK) inhibitor PKSI-527 attenuated arthritis signs and renal damage, and inhibited BK, PK, B1R and B2R expressions. The protein expressions of P38, p-P38 and p-JNK and IFN-gamma and TNF-alpha were inhibited by PKSI-527. Conclusions: These findings demonstrate that plasma KKS activation contributed to the renal injury of AA rats through MAPK signaling pathway. Plasma KKS might be a potential target for RA therapy.

Trichloroethylene (TCE) is able to induce trichloroethylene hypersensitivity syndrome (THS) with multi-system immune injuries. In our previous study, we found kallikrein-kinin system (KKS) activation, including the bradykinin B1 receptor (B1R), which contributed to immune organ injury in TCE sensitized mice. However, the mechanism of B1R mediating immune dysfunction is not clarified. The present study initiates to investigate the potential mechanism of B1R on liver injury. We establish a TCE sensitized BALB/c mouse model to explore the mechanism with or without a B1R inhibitor R715. We found B1R expression was increased in TCE sensitization-positive mice. As expect, hepatocyte intracellular organelles and mitochondria disappeared, glycogen particles reduced significantly as well in TCE sensitization-positive mice via the transmission electron microscopic examination, meanwhile, R715 alleviated the deteriorate above. The blockade of B1R resulted in a significant decreased p-ERK1/2 and increased p-AKT expression. The expression of CD68 kupffer cell and its relative cytokine, including IL-6 and TNF-α, increased in TCE sensitization-positive mice and decreased in R715 pretreatment TCE sensitization-positive mice. Together, the results demonstrate B1R plays a key role in ERK/MAPK and PI3K/AKT signal pathway activation and inflammation cytokine expression in immune liver injury induced by TCE. B1R exerts a pivotal role in the development of TCE induced liver injury.

Trichloroethylene (TCE) is known to induce skin disorders and multi-system dysfunction, but the mechanism of this multi-organ injury is not entirely clear. It was shown in a previous study that levels of pivotal end-products of the kallikrein-kinin system (KKS), i.e. bradykinin (BK) and BK receptors B1R/B2R, in the kidneys were increased by TCE exposure. Unfortunately, how BK and its receptors acted in the etiology of the induced renal injury is not clear. Thus, this study explored any correlation between BK receptors and immune renal injury in TCE-sensitized mice by blocking the BK receptors B1R/B2R. BALB/c mice were sensitized (via skin) by TCE, with or without pre-treatment with a B1R or B2R antagonist. Renal lesions, increased expressions of B1R, B2R, Kim-1, Lipocalin-2, and NF-κB p65 subunit on tubular epithelial cells were all observed in TCE-sensitized mice. Serum levels of creatinine (Cr), microglobulin α1 and β2, along with mRNA levels for inflammatory cytokines and NF- κB p65 in kidneys, were all increased by 72 h after a final challenge. Highly selective antagonist pre-treatment blocked B2R and significantly attenuated TCE-induced changes. Blocking B1R or B2R attenuated release of pro-inflammatory cytokines and activation of NF-κB signaling pathway (as reflected in lower up-regulation of pIκB and nuclear NF-κB p65 subunit, and down-regulation of IκB in the kidneys. These results provided evidence that TCE-sensitization caused KKS activation and enhanced the expression of B1R and B2R on tubular epithelial cells. This, in turn, accelerated NF-κB signaling pathway activation and amplified inflammatory cytokine release, which all likely contributed to TCE-induced immune renal injury.

Both NF-κB pathway and complement activation appear to be involved in kidney damage induced by trichloroethylene (TCE). However, any relationship between these two systems has not yet been established. The present study aimed to clarify the role of NF-κB in complement activation and renal injury in TCE-sensitized BALB/c mice. Mice were sensitized by an initial subcutaneous injection and repeated focal applications of TCE to dorsal skin at specified timepoints. NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) was injected (intraperitoneal) before the final two focal TCE challenges. In the experiments, mice had their blood and kidneys collected. Kidney function was evaluated via blood urea nitrogen (BUN) and creatinine (Cr) content; renal histology was examined using transmission electron microscopy (TEM). Kidney levels of phospho-p65 were assessed by Western blot and kidney mRNA levels of interleukin (IL)-1β, IL-6, IL-17, tumor necrosis factor (TNF)-α, and p65 by real-time quantitative PCR. Presence of C3 and C5b-9 membrane attack complexes in the kidneys was evaluated via immunohistochemistry. The results showed there was significant swelling, vacuolar degeneration in mitochondria, shrinkage of microvilli, disappearance of brush borders, segmental foot process fusion, and glomerular basement membrane thickening (or disrobing) in kidneys from TCE-sensitized mice. In conjunction with these changes, serum BUN and Cr levels were increased and IL-1β, IL-6, IL-17, and TNFα mRNA levels were elevated. Levels of p65 and phospho-p65 protein were also up-regulated, and there was significant C3 and C5b-9 deposition. PDTC pretreatment attenuated TCE-induced up-regulation of p65 and its phosphorylation, complement deposition, cytokine release, and renal damage. These results provide the first evidence that NF-κB pathway has an important role in TCE-induced renal damage mediated by enhanced complement activation in situ.