Genome-Wide CRISPR Screen Identifies an NF2-Adherens Junction Mechanistic Dependency for Cardiac Lineage.

BACKGROUND: Cardiomyocyte differentiation involves a stepwise clearance of repressors and fate-restricting regulators through the modulation of BMP (bone morphogenic protein)/Wnt-signaling pathways. However, the mechanisms and how regulatory roadblocks are removed with specific developmental signaling pathways remain unclear. METHODS: We conducted a genome-wide CRISPR screen to uncover essential regulators of cardiomyocyte specification in human embryonic stem cells using a myosin heavy chain 6 (MYH6)-GFP (green fluorescence protein) reporter system. After an independent secondary single guide ribonucleic acid validation of 25 candidates, we identified NF2 (neurofibromin 2), a moesin-ezrin-radixin like (MERLIN) tumor suppressor, as an upstream driver of early cardiomyocyte lineage specification. Independent monoclonal NF2 knockouts were generated using CRISPR-Cas9, and cell states were inferred through bulk RNA sequencing and protein expression analysis across differentiation time points. Terminal lineage differentiation was assessed by using an in vitro 2-dimensional-micropatterned gastruloid model, trilineage differentiation, and cardiomyocyte differentiation. Protein interaction and post-translation modification of NF2 with its interacting partners were assessed using site-directed mutagenesis, coimmunoprecipitation, and proximity ligation assays. RESULTS: Transcriptional regulation and trajectory inference from NF2-null cells reveal the loss of cardiomyocyte identity and the acquisition of nonmesodermal identity. Sustained elevation of early mesoderm lineage repressor SOX2 and upregulation of late anticardiac regulators CDX2 and MSX1 in NF2 knockout cells reflect a necessary role for NF2 in removing regulatory roadblocks. Furthermore, we found that NF2 and AMOT (angiomotin) cooperatively bind to YAP (yes-associated protein) during mesendoderm formation, thereby preventing YAP activation, independent of canonical MST (mammalian sterile 20-like serine-threonine protein kinase)-LATS (large tumor suppressor serine-threonine protein kinase) signaling. Mechanistically, cardiomyocyte lineage identity was rescued by wild-type and NF2 serine-518 phosphomutants, but not NF2 FERM (ezrin-radixin-meosin homology protein) domain blue-box mutants, demonstrating that the critical FERM domain-dependent formation of the AMOT-NF2-YAP scaffold complex at the adherens junction is required for early cardiomyocyte lineage differentiation. CONCLUSIONS: These results provide mechanistic insight into the essential role of NF2 during early epithelial-mesenchymal transition by sequestering the repressive effect of YAP and relieving regulatory roadblocks en route to cardiomyocytes.

Identification of key pathways and genes in the progression of silicosis based on WGCNA.

Silicosis, induced by inhaling silica particles in workplaces, is one of the most common occupational diseases. The prognosis of silicosis and its consequent fibrosis is extremely poor due to limited treatment modalities and lack of understanding of the disease mechanisms. In this study, a Wistar rat model for silicosis fibrosis was established by intratracheal instillation of silica (0, 50, 100 and 200 mg/mL, 1 mL) with the evidence of Hematoxylin and Eosin (HE) and Masson staining and the expressions of inflammatory and fibrotic proteins of rats' lung tissues. RNA of lung tissues of rats exposed to 200 mg/mL silica particles and normal saline for 14 d and 28 d was extracted and sequenced to detect differentially expressed genes (DEGs) and to identify silicosis fibrosis-associated modules and hub genes by Weighted gene co-expression network analysis (WGCNA). Predictions of gene functions and signaling pathways were conducted using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. In this study, it has been demonstrated the promising role of the Hippo signaling pathway in silicosis fibrosis, which will be conducive to elucidating the specific mechanism of pulmonary fibrosis induced by silica and to determining molecular initiating event (MIE) and adverse outcome pathway (AOP) of silicosis fibrosis.

Biomarkers of exposure and effect in the serum and urine of rats or workers exposed to 1-bromopropane.

Extensively used in several industries in China as a cleaning agent, 1-bromopropane (1-BP) has significant adverse effects on the central nervous system. However, neither its mechanism of action nor sensitive biomarkers related to it have been determined thus far. In this study, animal experiments and occupational surveys were performed to explore the typical exposure and effect biomarkers of neurotoxicity induced by 1-BP. Male Wistar rats were exposed to 0, 500, or 1000 ppm of 1-BP followed by pathological and biomarker analyses. An epidemiological survey was conducted on 71 workers each from 1-BP exposed and control groups. Serum and urine samples were collected for biomarker testing. cNSE represents neuron-specific enolase (NSE) in the cerebral cortex, where as sNSE represents NSE in the serum; similar terminology applies to S-100ß, and cyclooxygenase-2 (COX-2). In rats exposed to 1000 ppm 1-BP, pathological changes were observed in Purkinje cells, lumbar gray matter, and tibiofibular nerve, while levels of cNSE, cS-100ß, cCOX-2, sS-100ß, and sCOX-2 were significantly elevated at different time checkpoints. In the 500 ppm group, cCOX-2, sNSE, and sCOX-2 levels were significantly elevated at different time checkpoints. 1-BP and N-acetyl-S-(n-propyl)-L-cysteine (AcPrCys) were detected in rat urine, and there was a correlation between the level of sNSE or sCOX-2 and AcPrCys in the 500 ppm group. In the occupational epidemiological study, a significant correlation between AcPrCys and exposure concentration was also detected. The findings of this study indicated that AcPrCys was a sensitive exposure biomarker of 1-BP in rats as well as occupational populations.

A novel anatomical self-locking plate fixation for both-column acetabular fractures.

PURPOSE: To compare the stability of the posterior anatomic self-locking plate (PASP) with two types of popular reconstruction plate fixation, i.e. double reconstruction plate (DRP) and cross reconstruction plate (CRP), and to explore the influence of sitting and turning right/left on implants. METHODS: PASP, DRP and CRP were assembled on a finite element model of both-column fractures of the left acetabulum. A load of 600 N and a torque of 8 N·m were loaded on the S1 vertebral body to detect the change of stress and displacement when sitting and turning right/left. RESULTS: The peak stress and displacement of the three kinds of fixation methods under all loading conditions were CRP > DRP > PASP. The peak stress and displacement of PASP are 313.5 MPa and 1.15 mm respectively when turning right; and the minimal was 234.0 Mpa and 0.619 mm when turning left. CONCLUSION: PASP can provide higher stability than DRP and CRP for both-column acetabular fractures. The rational movement after posterior DRP and PASP fixation for acetabular fracture is to turn to the ipsilateral side, which can avoid implant failure.

[Renal damage of indium sulfate on rats].

OBJECTIVE: To study the renal damage of indium sulfate. METHODS: 32 healthy Wistar rats were randomly and equally divided into 3 dose groups( 52. 3 mg/kg、104. 6 mg/kg 261. 4 mg/kg) and one negative control group. Indium sulfate were orally given once a day successively 5 days a week for 8 weeks. Each group of rats was collected24 hour urine after the end of the posion. We tested the content of Cr, BUN, T-AOC, ALB in serum and the GSH activity in kidney by kids and detected the ß2-MG content in serum and urine by ELISA test. Inductively coupled plasma mass spectrometry( ICP-MS)method was used to detect the content of indium in whole blood, urine and kidney tissue of rats. Hematoxylin and eosin( H&E) staining was used to detect histological changes. RESULTS: During the experiment, all the rats were normal in activities, feed and drinking water, and they developed stably. In the period of seventh weeks and eighth weeks, the body weight of rats in high dose group was significantly lower than the control group( P <0. 05). Compared with the control group[( 1. 27 ± 0. 55), ( 0. 40 ± 0. 01) and( 0. 30 ±0. 06) µg/L], 3 dose group of indium in blood[( 44. 10 ± 23. 10), ( 52. 08 ± 21. 03) and( 67. 42 ± 45. 98) µg/L], urine[( 0. 72 ± 0. 13), ( 2. 75 ± 0. 15) and( 4. 31 ± 0. 33)µg/L]and kidney [( 1. 36 ± 0. 83), ( 1. 52 ± 0. 49) and( 2. 87 ± 0. 20) µg/L] were significantly increased( P < 0. 05). The level of Cr in serum in the high dose group were significantly higher than that in the control group [( 66. 06 ± 18. 62) and( 46. 53 ± 7. 95)µmol/L, P < 0. 05], the serum BUN content[( 3. 98 ± 0. 82) and( 4. 09 ± 0. 71) mmol/L] in the high dose group and middle dose group were significantly lower than the control group [( 4. 77 ± 0. 49) mmol/L, P < 0. 05]. Compared with the control group, 3 dose group of the ß2-MG in serum and urine were significantly increased( P < 0. 05), and the level of T-AOC[( 4. 87 ± 2. 36), ( 4. 50 ± 2. 33) and( 4. 00 ± 3. 29) U/m L] in serum and GSH[( 6. 41 ± 1. 86), ( 5. 06 ± 2. 09) and( 2. 77 ± 2. 64) µmol/( g prot) ] in renal tissue were significantly decreased[( 15. 20 ± 5. 43) U/m L and( 14. 74 ± 6. 47) µmol/( g prot), P < 0. 05]. Compared with the control group, the middle and high dose exposure group had significant inflammatory pathological changes, mainly manifested as glomerular swelling, renal tubular structure abnormalities and inflammatory cell infiltration. CONCLUSION: Indium sulfate can cause the accumulation of indium in the kidney, oxidative damage, pathological changes and dysfunction in the kidney of rats.

[Tumor markers of workers exposed to vinyl chloride].

OBJECTIVE: To detect the changes of serum tumor markers of vinyl chloride, and find the influencing factors of tumor markers. METHODS: Two hundred and twenty-three workers exposed to vinyl chloride from a chlor alkali plant and one hundred and forty-nine workers without occupational exposure to vinyl chloride were recruited into this study. Detected 11 tumor markers in serum of the objective. RESULTS: The contentsof carcino-embryonic antigen( CEA), alpha-fetoprotein( AFP), CA-199 and CA72-4 in exposed group increased with the length of service, while the content of neuron-specific enolase( NSE) decreased with the length of service, but there was no correlation between contents of tumor markers and the length of service, which had no statistical significance( P > 0. 05). By univariate analysis, the difference between the exposed group and the control group in the level of thiodiglycolic acid had statistical significance( Z =-16. 178, P < 0. 001). By univariate analysis, gender had effect on CEA, AFP and NSE( Z =-4. 815, -2. 052 and-4. 535, P < 0. 05), smoking had effect on CA-199( Z =-2. 016, P < 0. 05), vinyl chloride exposure had effect on AFP and NSE( Z =-3. 763 and-2. 140, P < 0. 05). By multivariate analysis, CEA and NSE of women were lower than those of men( t =-3. 696 and-5. 722, P < 0. 05). That vinyl chloride exposure was a factor in NSE and NSE of the exposed group were higher than the control group( t =2. 061, P < 0. 05). CONCLUSION: Under the current exposure concentration, exposure to vinyl chloride can change the contents of tumor markers. As the time of exposure to harmful factors increases, the level of tumor markers changes, and the possibility of tumor increases. The level of thiodiglycolic acid may be related to some tumor markers. The influencing factors of different tumor markers are different.

[Research on olaquindox induced endoplasmic reticulum stress related apoptosis on nephrotoxicity].

OBJECTIVE: Renal tubular epithelial cell were exposed to olaquindox and detected the ROS and apoptosis related proteins, to investigate the renal tubular epithelial cell apoptosis through endoplasmic reticulum stress mediated pathway induced by olaquindox. METHODS: MTT assay (1, 2, 3, 4, 5, 6, 7and 8 µmol/ml olaquindox exposure) was used to detect the effects of olaquindox on renal tubular epithelial cell proliferation to determine test concentrations. Hoechst-33258 was used to detect morphological changes on apoptotic cells in each group. Flow cytometry method was applied to detect the apoptosis rate and intracellular reactive oxygen, and western blot assay was performed to detect the levels of endoplasmic reticulum stress-related apoptosis proteins, GRP78, GRP94 and CHOP. RESULTS: According to results of the MTT test, 1, 2, 3 and 4 µmol/ml olaquindox concentrations were determined for apoptosis analysis. With the increase of olaquindox concentration, apoptosis rate and levels of endoplasmic reticulum stress related apoptosis pathway protein GRP78, GRP94 and CHOP increased, levels of ROS were increased in every groups (P < 0.05) in 2 µmol/ml olaquindox groups and above. With the prolongation of olaquindox exposure, apoptosis rate and levels of endoplasmic reticulum stress related apoptosis pathway protein GRP78 and GRP94 increased in 12 and 24 h olaquindox exposure groups, whereas in groups of olaquindox exposed for 6, 12 and 24 h, levels of ROS and endoplasmic reticulum stress related apoptosis pathway protein CHOP increased (P < 0.05). CONCLUSION: Olaquindox can induce renal tubular epithelial cells to apoptosis and cause the renal toxicity, and the endoplasmic reticulum stress-related apoptosis maybe the associated toxicity pathway.

Cloning and characterization of AabHLH1, a bHLH transcription factor that positively regulates artemisinin biosynthesis in Artemisia annua.

Amorpha-4,11-diene synthase (ADS) and Cyt P450 monooxygenase (CYP71AV1) in Artemisia annua L. are two key enzymes involved in the biosynthesis of artemisinin. The promoters of ADS and CYP71AV1 contain E-box elements, which are putative binding sites for basic helix-loop-helix (bHLH) transcription factors. This study successfully isolated a bHLH transcription factor gene from A. annua, designated as AabHLH1, from a cDNA library of the glandular secretory trichomes (GSTs) in which artemisinin is synthesized and sequestered. AabHLH1 encodes a protein of 650 amino acids containing one putative bHLH domain. AabHLH1 and ADS genes were strongly induced by ABA and the fungal elicitor, chitosan. The transient expression analysis of the AabHLH1-green fluorescent protein (GFP) reporter gene revealed that AabHLH1 was targeted to nuclei. Biochemical analysis demonstrated that the AabHLH1 protein was capable of binding to the E-box cis-elements, present in both ADS and CYP71AV1 promoters, and possessed transactivation activity in yeast. In addition, transient co-transformation of AabHLH1 and CYP71AV1Pro::GUS in A. annua leaves showed a significant activation of the expression of the GUS (ß-glucuronidase) gene in transformed A. annua, but mutation of the E-boxes resulted in abolition of activation, suggesting that the E-box is important for the CYP71AV1 promoter activity. Furthermore, transient expression of AabHLH1 in A. annua leaves increased transcript levels of the genes involved in artemisinin biosynthesis, such as ADS, CYP71AV1 and HMGR. These results suggest that AabHLH1 can positively regulate the biosynthesis of artemisinin.

[Establishment and application of an in vitro model for dynamic direct exposure of lung epithelial cells (A549) to airborne compounds].

OBJECTIVE: To establish and apply a dynamic in vitro model for direct exposure of human cells to gaseous contaminants to investigate the cellular responses to airborne chemical exposures. METHODS: Nitrogen dioxide (NO2) was selected as a model gas compound. Standard test atmospheres were generated (5.1 - 20.7 mg/m3), using clean air dilution method. A549 pulmonary type II-like epithelial cell lines and skin fibroblasts were grown on porous membranes. Human cells on transwell inserts were placed in horizontal diffusion chambers and exposed to various airborne concentrations of NO2 directly at the air- liquid interface for 1 h at 37 degrees C, 15 ml/min. The gasoline exhuast at different load (idle with cold start, 50% and 100% load) collected in Tedlar air bag directly exposed A549 cell for 15 min at 37 degrees C, 10 ml/min. Cytotoxicity of the test gas was investigated using the MTT, NRU (neutral red uptake) and CCK-8 assays. RESULTS: Dose-dependent effects of NO2 were observed in human cells tested which resulted in a significant reduction of cell viability at different concentrations of NO2 (MTT estimated curve:gamma(MTT) = 0.989 - 0.021 chi (NO(2)) R2 = 0.84, P < 0.01; NUR estimated curve:gamma(NUR) = 1.032 - 0.026 chi (NO(2)) R2 = 0.88, P < 0.01). Gasoline exhaust exposure at different load resulted in a decreased cell viability (significantly CCK-8 and MTT levels reduction than air exposure group). CONCLUSION: The dynamic direct exposure method can be used for in vitro inhalational and dermal toxicity studies and potentially as an new technology for biomonitoring of airborne contaminants in future occupational and environmental toxicity assessments.

[Study on expression of functional proteins related with Schwann cell of rats in acrylamide exposure and convalescent].

OBJECTIVE: To explore the changes of four functional proteins which are related to Schwan cells (SCs), including myelin-associated glycoprotein (MAG), nerve growth factor (NGF), p75 neurotrophin receptor (p75NTR) and neural cell adhesion molecule (NCAM) on damage and repair of peripheral nerve induced by acrylamide (Acr). From the changes of the protein level, some meaningful information for the mechanism of Acr neurotoxicity and the screening of biomarkers might be acquired. METHODS: Rats were administrated with Acr at dose of 7. 5, 15 and 20 mg/kg by intraperitoneal injection for 3 weeks, high-dose group were observed for 4 weeks after 3 weeks exposure of Acr to create an animal model of peripheral nerve in injury and repair. Protein level of MAG, p75NTR, NGF and NCAM in rat sciatic nerve at the end of exposure and convalescent were measured by western blot. The level of MAG in plasma at the end of exposure and convalescent was measured by ELISA. RESULTS: (1) Rats treated with Acr appeared peripheral nerve damage symptom and began to recover after 4 weeks. The abnormal symptoms in female group were heavier than that of males, especially the high dose group. (2) Compared with the control group, the level of MAG decreased in the medium dose group and high dose group (P < 0.05), the level of p75NTR increased in high dose group (P < 0.05). There were no significant changes in the level of NGF between the control group and treated groups of male rats. Compared with the male control group, the level of NCAM in the the high dose group increased (P < 0.05). (3) Compared with the control group, the level of plasma MAG in the high dose group decreased (P < 0.05), while that in the recovery group was slightly increased. CONCLUSION: The changes of those functional proteins may reflect the state of the peripheral nerve damage induced by Acr. The downregulation of MAG in rat plasma may be related with that in sciatic nerve.

[Effect of rich-D-transallethrin on amino acid neurotransmitters in rat brain].

OBJECTIVE: To discuss the effect of rich-D-transallethrin on amino acid neurotransmitters in rats central nerves system and pathological examination of brain tissues, hypophysis and sciatic nerve. METHODS: Ninety-six male and female Wistar rats were randomly divided into four groups according to body weight, which were exposed to rich-d-transallthrin aerosol at different dose (0, 9.6, 45.8, 166 mg/m3) for 6 h/d, 7 d/week for 28 consecutive days. Neurobehavior were observed, gait and grip strength were measured during exposure. At the end of treatment the rats in all groups were sacrificed. The content of glutamate (Glu) and glycine (Gly) in brain tissues were determined and the pathological examination of brain tissues, hypophysis and sciatic nerve were conducted. RESULTS: The grip strength in 166.0 mg/m3 exposure group was significantly decreased (P < 0.05) compared with control group. The level of glutamate and glycine in female rats brain tissues was also significantly reduced (P < 0.05) after treatment with rich-d-transallethrin aerosol for 4 weeks. The result of pathological examination showed that cerebral cortex, hippocampus and cerebellum appeared neuron degeneration and slight axon swelling and myelin sheath destruction in sciatic nerve induced by 166.0 mg/m3 rich-d-transallethrin aerosol. CONCLUSION: The changes of Glu, Gly and pathological examination could be related to treatment with rich-d-transallethrin, in the meanwhile the major effect on nervous system appeared to be the cerebral cortex, hippocampal neuron and peripheral motor nerve.

[Experimental study on adsorption effect of activated carbon to abamectin].

OBJECTIVE: To investigate the adsorbability of activated charcoals for abamectin in vitro at various pH and concentrations. METHODS: Three concentrations of abamectin solutions (10, 5 and 2.5 g/L) with different pH value ( pH1.9 and pH6.8, respectively) were mixed with activated charcoal and control, respectively. The concentrations of abamectin were measured by UV-visible spectrophotometry after vibrated, incubated and centrifuged on different time points. Calculate the residue rate of abamectin and adsorption rate of activated carbon for avermectins. RESULTS: The abamectin was significantly decreased in activated charcoal group compared with control group and the absorption ability of activated charcoal was higher on low concentration of abamectin. CONCLUSION: In vitro experiment showed that activated charcoal can significantly adsorb abamectin.

[Progress on treatment of acetabular quadrilateral plate injury].

Acetabular quadrilateral plate injury has become a hot spot and focus in the field of orthopaedic trauma and pelvic floor function in recent years. Although there are five fracture types,they are all based on fracture morphology,without considering the pulling force of ligaments,joint capsular and muscles. A perfect classification needs to describe the displacement of bone mass in three-dimensional space to better guide reduction and fixation. The seven incision and exposure methods are still the traditional open-eye surgery,and how to protect the criss-crossing vascular neural network and pelvic organs is still the focus. Quadrilateral defect causes dislocation of artificial hip joint,and quantitative evaluation of quadrilateral defect volume and revision techniques are still a hot topic. In this paper,the viewpoints of three-dimensional network structure of acetabular pelvic vascular anatomy,anatomical surgical target channel and fixation anchor point of acetabular fracture reduction are proposed to design new techniques for accurate and minimally invasive surgical operations,in order to realize the requirements of rapid orthopedic rehabilitation.

Hidden blood loss and the influential factors after minimally invasive treatment of posterior pelvic ring injury with sacroiliac screw.

BACKGROUND: To analyze the perioperative bleeding and hidden blood loss (HBL) of sacroiliac screw minimally invasive treatment of pelvic posterior ring injury and explore the influential factors of HBL after operation for providing reference for clinical treatment. METHOD: A retrospective analysis was conducted on data from 369 patients with posterior pelvic ring injuries treated with sacroiliac screws internal fixation at our hospital from January 2015 to January 2022. The research was registered in the Chinese Clinical Trial Registry in July 2022 (ChiCTR2200061866). The total blood loss (TBL) and HBL of patients were counted, and the factors such as gender, age, and surgical duration were statistically analyzed. The influential factors of HBL were analyzed by multiple linear regression. RESULTS: The TBL was 417.96 ± 98.05 ml, of which the visible blood loss (VBL) was 37.00 ± 9.0 ml and the HBL was 380.96 ± 68.8 ml. The HBL accounted for 91.14 ± 7.36% of the TBL. Gender, surgical duration, fixed position, and fixed depth had significant effects on the HBL (P < 0.05). CONCLUSIONS: The HBL was the main cause of anemia after minimally invasive treatment of posterior pelvic ring injury with a sacroiliac screw. Gender, surgical duration, fixed position, and fixed depth were closely related to the occurrence of HBL. In clinical treatment, we should consider these influential factors and take effective measures to reduce the impact of HBL on patients.

Chronic Lead Exposure in Adult Mice: Associations with miR-671/CDR1as Regulation, NF-κB Signaling, and Alzheimer's Disease-like Pathology.

Long-term exposure to lead (Pb) can result in chronic damage to the body through accumulation in the central nervous system (CNS) leading to neurodegenerative diseases, such as Alzheimer's disease (AD). This study delves into the intricate role of miR-671/CDR1as regulation in the etiology of AD-like lesions triggered by chronic Pb exposure in adult mice. To emulate the chronic effects of Pb, we established a rodent model spanning 10 months of controlled Pb administration, dividing 52 C57BL/6J mice into groups receiving varying concentrations of Pb (1, 2, or 4 g/L) alongside an unexposed control. Blood Pb levels were monitored using serum samples to ensure accurate dosing and to correlate with observed toxicological outcomes. Utilizing the Morris water maze, a robust behavioral assay for assessing cognitive functions, we documented a dose-dependent decline in learning and memory capabilities among the Pb-exposed mice. Histopathological examination of the hippocampal tissue revealed tell-tale signs of AD-like neurodegeneration, characterized by the accumulation of amyloid plaques and neurofibrillary tangles. At the molecular level, a significant upregulation of AD-associated genes, namely amyloid precursor protein (APP), ß-secretase 1 (BACE1), and tau, was observed in the hippocampal tissue of Pb-exposed mice. This was accompanied by a corresponding surge in the protein levels of APP, BACE1, amyloid-ß (Aß), and phosphorylated tau (p-tau), further implicating Pb in the dysregulation of these key AD markers. The expression of CDR1as, a long non-coding RNA implicated in AD pathogenesis, was found to be suppressed in Pb-exposed mice. This observation suggests a potential mechanistic link between Pb-induced neurotoxicity and the dysregulation of the CDR1as/miR-671 axis, which warrants further investigation. Moreover, our study identified a dose-dependent alteration in the intracellular and extracellular levels of the transcription factor nuclear factor-kappa B (NF-κB). This finding implicates Pb in the modulation of NF-κB signaling, a pathway that plays a pivotal role in neuroinflammation and neurodegeneration. In conclusion, our findings underscored the deleterious effects of Pb exposure on the CNS, leading to the development of AD-like pathology. The observed modulation of NF-κB signaling and miR-671/CDR1as regulation provides a plausible mechanistic framework for understanding the neurotoxic effects of Pb and its potential contribution to AD pathogenesis.

[Genetic damage in peripheral blood lymphocyte of 1,3-butadiene workers].

OBJECTIVE: To investigate the DNA and chromosome damage in peripheral blood lymphocyte of workers occupationally exposed to 1,3-butadiene (BD). METHODS: Personal information including occupational history, age, sex, smoking and drinking status was collected by the questionnaire. Gas chromatography was used to analyze the BD level. One hundred and eighty 1,3-butadiene workers and 58 controls without occupational BD exposure were investigated. Comet assay and cytokinesis-block micronucleus (CBMN) detection were used to evaluate DNA and chromosomal damage levels in peripheral blood lymphocyte. RESULTS: The concentration of BD in the working environment of BD-exposed workers was 1.80 (0.59-2.76) mg/m3. The rate of CBMN, NPB, NBUD and Olive TM of lymphocyte in BD-exposed workers [(6.76 +/- 4.99) per thousand, 1.00 (0.00-4.00), 2.00 (0.00-7.00) and 4.64 (3.50-5.98), respectively] were higher than those in controls [(3.10 +/- 2.65) per thousdand, 0.00 (0.00-2.80), 1.00 (0.00- 5.00) and 2.34 (0.82-3.93), P < 0.01]. According to the length of work, 180 BD-exposed workers were classified into 3 groups: 1 yrs-, 14.0 yrs- and 20.0 yrs-group, respectively after adjusting the age,sex, smoking and drinking, the rate of CBMN was a rising tendency along with the increase in length of work. CONCLUSION: Under present BD exposure levels, both comet assay and Cytokinesis-block micronucleus test could detect BD-induced genotoxicity in BD-exposed workers, and are more suitable to assess the cumulative damage effect on DNA.

Deleterious Mechanical Deformation Selects Mechanoresilient Cancer Cells with Enhanced Proliferation and Chemoresistance.

Cancer cells in secondary tumors are found to form metastases more efficiently as compared to their primary tumor counterparts. This is partially due to the unfavorable microenvironments encountered by metastasizing cancer cells that result in the survival of a more metastatic phenotype from the original population. However, the role of deleterious mechanical stresses in this change of metastatic potential is unclear. Here, by forcing cancer cells to flow through small capillary-sized constrictions, it is demonstrated that mechanical deformation can select a tumor cell subpopulation that exhibits resilience to mechanical squeezing-induced cell death. Transcriptomic profiling reveals up-regulated proliferation and DNA damage response pathways in this subpopulation, which are further translated into a more proliferative and chemotherapy-resistant phenotype. These results highlight a potential link between the microenvironmental physical stresses and the enhanced malignancy of metastasizing cancer cells which may be utilized as a therapeutic strategy in preventing the metastatic spread of cancer cells.

A Promising transwell co-culture cell model for silicosis.

Silicosis, one of the most ancient occupational diseases, has not been elucidated and addressed until now. Although some medicines were partly effective in the clinical treatment and certain plausible mechanisms were partially ascertained, it is still urgent and necessary to explore its real and specific pathogenesis. With the development of the cell co-culture technique, the transwell co-culture system has been applied in several scientific fields, which is able to simulate the pathogenic process in vivo to the largest extent. In this sense, a transwell co-culture cell model for silicosis including RFL-6 (lung fibroblasts of rats), RLE-6TN (type II alveolar epithelial cells of rats) and NR8383 (alveolar macrophages of rats) cells was successfully established and the developmental process of silicosis in this model could be roughly divided into three stages: inflammatory stage, progressive stage and fibrotic stage, as evidenced by the morphological features and the inflammatory cytokines and fibrotic proteins of supernatants and the cells in this system. Results of this project will pave the way for clarifying the mechanism of silicosis and building a practical platform for the further exploration of molecular initiating event (MIE) and adverse outcome pathway (AOP) of silicosis.

Current status of robot-assisted surgery in the clinical application of trauma orthopedics in China: A systematic review.

Background and Aims: To elaborate on the development and characteristics of trauma orthopedic robots and their real curative effect in a clinical application through the collection and analysis of relevant literature and reported clinical results. Method: We conducted the Embase, ScienceDirect, Pubmed, Medline, Wanfang, CNKI, and VIP search of the literature on robotic-assisted surgery in trauma orthopedics in China. We combined search terms with "robotic surgery/artificial intelligence surgery/navigation surgery," "trauma/trauma orthopedics," and "China/Chinese." The exclusion criteria were: (1) articles in languages other than English or Chinese, (2) articles focused on other topics other than robotic-assisted surgery in trauma orthopedics of China, (3) article types were not clinical studies (reviews, basic research, etc.), and (4) articles were not included in the Chinese core journals or science citation index. Authors, type of surgery, robot type, and clinical research results were recorded and analyzed. Results: There were three categories of surgical robots in the clinical application of trauma orthopedics (TiRobot, electromagnetic navigation surgical robots, and small medical robots developed by Beijing Jishuitan Hospital). In terms of blood loss, the fluoroscopy time, and fluoroscopy frequency, most studies found that the robot group was significantly better than the traditional group. Conclusions: Robot-assisted surgery has obvious advantages in accuracy, stability, and reducing intraoperative radiation exposure, but there is no final conclusion about functional recovery.

Acrylamide-induced damage to postsynaptic plasticity is CYP2E1 dependent in an SH-SY5Y co-culture system.

Acrylamide (ACR), a neurotoxic substance, is characterized by a range of industrial and population exposures. The effects of ACR on synapses have been examined, but the regulation and molecular mechanism of key proteins related to ACR and its metabolite glycidamide (GA) have not been elucidated. In this study, we constructed two co-culture systems to mimic neurons that do not express and overexpress CYP2E1. In these co-cultures, we observed the effects and relative influence of ACR and GA on cell survival as well as synaptic structural and functional plasticity. Next, we investigated the relationship between ACR-induced nerve damage and key proteins in the postsynaptic membrane. After ACR exposure, cell death and synaptic damage were significantly worse in CYP2E1-overexpressing co-culture systems, suggesting that ACR-induced neurotoxicity may be related to metabolic efficiency (including CYP2E1 activity). Moreover, with increasing doses of ACR, the key postsynaptic membrane proteins PSD-95 expression was reduced and CaMKII and NMDAR-2B phosphorylation was increased. ACR exposure also triggered a rapid dose- and time-dependent increase in intracellular Ca2+, whose changes can affect the expression of the above-mentioned key proteins. In summary, we clarified the relationship between ACR exposure, neuronal damage and postsynaptic plasticity and proposed an ACR-CYP2E1-GA: Ca2+-PSD-95-NMDAR-Ca2+-CaMKII effect chain. This information will further improve the development of an alternative pathway strategy for investigating the risk posed by ACR.