ObjectiveTo construct a stable monoclonal human embryonic kidney 293 （HEK293） cell line expressing recombinant human coagulation factor Ⅶ （rhFⅦ） and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells， and monoclonal stable transfected cell lines were selected using geneticin （G418）. The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction （RT-PCR）. The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay （ELISA）， and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence， indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8， and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was （1.27±0.09） mg/L， and the highest procoagulant activity was （380.29±13.80）%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.

BACKGROUND: Tissue engineering holds promise for vascular repair and regeneration by mimicking the extracellular matrix of blood vessels. However, achieving a functional and thick vascular wall with aligned fiber architecture by electrospinning remains a significant challenge. METHODS: A novel electrospinning setup was developed that utilizes an auxiliary electrode and a spring. The impact of process parameters on fiber size and morphology was investigated. The structure and functions of the scaffolds were evaluated through material characterization and assessments of cellular biocompatibility. RESULTS: The new setup enabled controlled deposition of fibers in different designed orientations. The fabricated small-diameter vascular scaffolds consisted of an inner layer of longitudinally oriented fibers and an outer layer of circumferentially oriented fibers (L + C vascular scaffold). Key parameters, including rotational speed, the utilization of the auxiliary electrode, and top-to-collector distance (TCD) significantly influenced fiber orientation. Additionally, voltage, TCD, feed rate, needle size, auxiliary electrode and collector-auxiliary electrode distance affected fiber diameter and distribution. Mechanical advantages and improved surface wettability of L + C vascular scaffold were confirmed through tensile testing and water contact angle. Cellular experiments indicated that L + C vascular scaffold facilitated cell adhesion and proliferation, with human umbilical vein endothelial cells and smooth muscle cells attaching and elongating along the fiber direction of the inner and outer layer, respectively. CONCLUSION This study demonstrated the feasibility of fabricating fiber-aligned, thick-walled vascular scaffolds using a modified electrospinning setup. The findings provided insights into how the auxiliary electrode, specific collector influenced fiber deposition, potentially advancing biomimetic vascular scaffold engineering.

Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

BACKGROUND: Tissue engineering holds promise for vascular repair and regeneration by mimicking the extracellular matrix of blood vessels. However, achieving a functional and thick vascular wall with aligned fiber architecture by electrospinning remains a significant challenge. METHODS: A novel electrospinning setup was developed that utilizes an auxiliary electrode and a spring. The impact of process parameters on fiber size and morphology was investigated. The structure and functions of the scaffolds were evaluated through material characterization and assessments of cellular biocompatibility. RESULTS: The new setup enabled controlled deposition of fibers in different designed orientations. The fabricated small-diameter vascular scaffolds consisted of an inner layer of longitudinally oriented fibers and an outer layer of circumferentially oriented fibers (L + C vascular scaffold). Key parameters, including rotational speed, the utilization of the auxiliary electrode, and top-to-collector distance (TCD) significantly influenced fiber orientation. Additionally, voltage, TCD, feed rate, needle size, auxiliary electrode and collector-auxiliary electrode distance affected fiber diameter and distribution. Mechanical advantages and improved surface wettability of L + C vascular scaffold were confirmed through tensile testing and water contact angle. Cellular experiments indicated that L + C vascular scaffold facilitated cell adhesion and proliferation, with human umbilical vein endothelial cells and smooth muscle cells attaching and elongating along the fiber direction of the inner and outer layer, respectively. CONCLUSION This study demonstrated the feasibility of fabricating fiber-aligned, thick-walled vascular scaffolds using a modified electrospinning setup. The findings provided insights into how the auxiliary electrode, specific collector influenced fiber deposition, potentially advancing biomimetic vascular scaffold engineering.

Nanomaterials have been used in a variety of industries recently,involving foods,chemi-cals and biomedicine.There are multiple routes through which humans are exposed to nanomaterials,and their toxic effects deserve more attention.In vivo studies have confirmed that nanomaterials expo-sure can lead to toxicity in such target organs as the heart,liver,kidney,skin and nerve.The toxicity mechanism is related to changes in organelles such as the endoplasmic reticulum,lysosomes and mito-chondria.Increasing studies suggest that mitochondria are critical targets for the toxicity of nanomaterials.Mitochondrial biogenesis serves as an important mechanism for maintaining mitochondrial homeostasis,which plays a vital role in the process of nanomaterials-induced cellular toxicity.This article summarizes the current research on the effects of nanomaterials on mitochondrial biogenesis,and elaborates the mechanism through which nanomaterials disrupt mitochondrial biogenesis by triggering oxidative stress,upsetting the homeostasis of calcium ion and disturbing toxicity pathways.This article is expected to provide a reference for toxicity testing and risk assessment of nanomaterials.

Objective To understand the instrument distribution and capacities of inter-laboratory comparisons among occupational medical examination (OME) institutions in Guangdong Province. Methods A total of 291 OME institutions in Guangdong Province that were within the validity period of filing in June 2024 and 101 OME institutions that participated in inter-laboratory comparisons in 2023 were selected as the study subjects. Their filing categories, instrument distribution and external quality assessment in laboratories were collected and analyzed. Results The location of 291 OME institutions distributed in 21 prefectural-level cities in the province, mainly in the Pearl River Delta (PRD) region, accounting for 68.4% of the total. The public hospitals, private medical institutions, the center for disease control and prevention and occupational disease prevention and treatment institutions (hereinafter referred to as "occupational prevention institutions") accounted for 61.2%, 28.5% and 10.3%, respectively. The filing rates for physical factors, chemical factors, and dust categories among OME institutions were 91.1%, 86.9%, and 86.6%, respectively. The average number of institutional instruments, from high to low, was 53.8, 38.8 and 38.2 per unit in occupational prevention institutions, private medical institutions and public hospitals, respectively. The passing rates of inter-laboratory comparisons among OME institutions of external quality assessment in laboratories among OME institutes in Guangdong Province, from high to low were occupational prevention institutions, public hospitals and private medical institutions (96.2% vs 86.8% vs 73.0%, P<0.05). Conclusion The regional distribution of OME institutes in Guangdong Province is unbalanced, mainly concentrated in the PRD region. Low overall level of OME capacity is found in public hospitals and private medical institutions. Leading by the occupational prevention institutions, public hospitals and private medical institutions should strengthen the development and investment in OME hardware, in order to improve the OME level.

Objective To analyze the status and its influencing factors of high-frequency hearing loss (HFHL) among occupational noise-exposed workers. Methods A total of 6 764 noise-exposed workers who participated in occupational medical examinations during their employment were selected as the study subjects using the judgmental sampling method. Their pure-tone audiometry results were analyzed. Results The detection rate of HFHL among the study subjects was 10.3%. Binary logistic regression analysis result revealed that males had a higher risk of HFHL than females (P<0.05). The HFHL risk of workers increased with age (all P<0.01). Workers with longer noise exposure durations had a higher risk of HFHL (all P<0.01). Smokers had a higher detection rate of HFHL than non-smokers (P<0.01). Workers in the mining industry, electricity/heat/gas and water production and supply industry, and other industries had a higher HFHL detection rate compared with manufacturing workers (all P<0.05). Medium-sized enterprise workers had a lower HFHL detection rate compared with micro-sized enterprise workers (P<0.05), while workers in private enterprises had a higher HFHL detection rate compared with those in state-owned enterprises (P<0.05). Workers who exposed to noise combined with dust, lead, high temperature, or multiple factors had a higher HFHL risk than those only exposed to noise (all P<0.05). Conclusion Gender, age, noise exposure duration, smoking, industry, and exposure to noise combined with other occupational hazard factors are all influencing factors for HFHL among noise-exposed workers. Targeted occupational health education on noise hazard prevention and occupational medical surveillance should be strengthened for key populations in priority industries, including male workers, older employees, individuals with prolonged noise exposure, smokers, and those concurrently exposed to noise and one of the following: dust, lead, high temperatures, or multiple occupational factors.

The lung function of patients with occupational pneumoconiosis (hereinafter referred to as "pneumoconiosis") combined with chronic obstructive pulmonary disease (COPD) declines rapidly. Inhaled pharmacotherapy is crucial in the treatment of patients with pneumoconiosis combined with COPD. At present, pressurized metered-dose inhalers, dry powder inhalers, and soft mist inhalers are commonly used to deliver bronchodilators (including long-acting β2-adrenergic agonists, long-acting muscarinic antagonists, or their fixed-dose combinations) in inhaled pharmacotherapy. Peak inspiratory flow (PIF) is a commonly used indicator for evaluating the effectiveness of inhalation therapy. Patients with good coordination of hand and mouth, and a high PIF (≥30 L/min) can be treated with pressurized metered-dose inhalers or dry powder inhalers, whereas patients with poor coordination or insufficient PIF (<30 L/min) should be treated with pressurized metered-dose inhalers combined with a spacer or with nebulizers. In addition, the effectiveness of inhalation therapy is affected by individual factors, such as treatment adherence and pulmonary function, as well as drug-related factors, including the use of inhaled corticosteroids and the selection of bronchodilators. Inhaled pharmacotherapy appears to have substantial potential in the treatment of pneumoconiosis complicated with COPD. Future efforts should focus on the development of more effective inhaled drugs with fewer adverse effects and more user-friendly inhalation devices, as well as on optimizing inhalation treatment regimens to enable precision therapy and improve therapeutic efficacy.

Objective To analyze the occupational health literacy (OHL) level and its influencing factors of workers in non-metallic mineral product industry in Yunfu City. Methods A total of 947 frontline workers from 24 non-metallic mineral products enterprises in Yunfu City were selected as the research subjects using the stratified random sampling method. The OHL level of the workers were assessed using the Occupational Health Literacy Questionnaire of National Key Populations. Results The overall OHL level of the research subjects was 58.3% (552/947). The OHL levels across four dimensions, from highest to lowest, were basic knowledge of occupational health protection (94.7%), healthy work practices and behaviors (81.8%), legal knowledge of occupational health (65.5%), and basic skills of occupational health protection (25.9%). The results of binary logistic regression analysis showed that workers with 2.0-<10.0 years and ≥10 years of work experience had higher OHL levels than those with <2.0 years of work experience (all P<0.01). Workers with a high school education or above had higher OHL levels than those with a junior high school education or below (all P<0.01). Workers in large- and medium-sized enterprises had higher OHL levels than those in small and micro-sized enterprises (both P<0.01). Conclusion The OHL levels of workers in Yunfu City's non-metallic mineral products industry can be further improved, particularly the occupational health protection skills and related legal knowledge. Workers with short seniority, low educational level, and in small and micro enterprises should be the key groups for improving OHL levels.

Objective To analyze a case of violations by an occupational medical examination (OME) institution and to explore the key control points for the supervision and management of OME institutions, as well as the core role of quality assessment in this context. Methods An OME institution suspected of illegal activities was used as the study subject. Retrospective analysis was conducted. Clues of suspected violations were identified by an on-site quality assessment. After investigation and verification by the local health authorities, legal action was taken against the institution for its violations. Results During an on-site quality assessment, the Guangdong Province OME quality control expert group discovered that the OME institution violated regulations, including unqualified personnel file, exceeding the scope of services category, issuing false reports, failing to report suspected occupational diseases on time, and failing to notify workers about suspected occupational diseases as required. The evidence was then submitted to the Guangdong Province OME Quality Control Center, which subsequently forwarded the case to local health administration department for filing and investigation. After the investigation, penalties were imposed on the OME institution for its illegal activities. Conclusion The key supervision and inspection points in the quality assessment of OME institutions include personnel file configuration, the quality control management system and its implementation, the quality of OME reports, and information reporting. Quality assessment plays a pivotal role in ensuring the legal and compliant practice of OME institutions, safeguarding the health rights and interests of workers, and enhancing the overall standard of the OME industry.