1.Mechanism of Yishen Tongluo Formula regulating the TLR4/MyD88/NF-κB signaling pathway to ameliorate pyroptosis in diabetic nephropathy mice
Yifei ZHANG ; Zijing CAO ; Zeyu ZHANG ; Xuehui BAI ; Jingyi TANG ; Junyu XI ; Jiayi WANG ; Yiran XIE ; Yuqi WU ; Xi GUO ; Zhongjie LIU ; Weijing LIU
Journal of Beijing University of Traditional Chinese Medicine 2025;48(1):21-33
Objective:
To investigate the mechanism of Yishen Tongluo Formula in ameliorating renal pyroptosis in diabetic nephropathy mice by regulating the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway.
Methods:
Sixty C57BL/6 male mice were randomly divided into control (10 mice) and intervention groups (50 mice) using random number table method. The diabetes nephropathy model was established by intraperitoneally injecting streptozotocin(50 mg/kg). After modeling, the intervention group was further divided into model, semaglutide (40 μg/kg), and high-, medium-, and low-dose Yishen Tongluo Formula groups (15.6, 7.8, and 3.9 g/kg, respectively) using random number table method. The high-, medium-, and low-dose Yishen Tongluo Formula groups were administered corresponding doses of medication by gavage, the semaglutide group received a subcutaneous injection of semaglutide injection, and the control group and model groups were administered distilled water by gavage for 12 consecutive weeks. Random blood glucose levels of mice in each group were monitored, and the 24-h urinary protein content was measured using biochemical method every 4 weeks; after treatment, the serum creatinine and urea nitrogen levels were measured using biochemical method. The weight of the kidneys was measured, and the renal index was calculated. Hematoxylin and eosin, periodic acid-Schiff, periodic Schiff-methenamine, and Masson staining were used to observe the pathological changes in renal tissue. An enzyme-linked immunosorbent assay was used to detect urinary β2-microglobulin (β2-MG), neutrophil gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1) levels. Western blotting and real-time fluorescence PCR were used to detect the relative protein and mRNA expression levels of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3), Caspase-1, gasdermin D (GSDMD), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in renal tissue. Immunohistochemistry was used to detect the proportion of protein staining area of the TLR4, MyD88, and NF-κB in renal tissue.
Results:
Compared with the control group, the random blood glucose, 24-h urinary protein, serum creatinine, urea nitrogen, and renal index of the model group increased, and the urine β2-MG, NGAL, and KIM-1 levels increased. The relative protein and mRNA expression levels of NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 in renal tissue increased, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas increased (P<0.05). Pathological changes such as glomerular hypertrophy were observed in the renal tissue of the model group. Compared with the model group, the Yishen Tongluo Formula high-dose group showed a decrease in random blood glucose after 12 weeks of treatment (P<0.05). The Yishen Tongluo Formula high- and medium-dose groups showed a decrease in 24-h urinary protein, creatinine, urea nitrogen, and renal index, as well as decreased β2-MG, NGAL, and KIM-1 levels. NLRP3, Caspase-1, GSDMD, IL-1 β, and IL-18 relative protein and mRNA expression levels were also reduced, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas was reduced (P<0.05). Pathological damage to renal tissue was ameliorated.
Conclusion
Yishen Tongluo Formula may exert protective renal effects by inhibiting renal pyroptosis and alleviating tubular interstitial injury in diabetic nephropathy mice by regulating the TLR4/MyD88/NF-κB signaling pathway.
2.Yishen Tongluo Prescription Ameliorates Oxidative Stress Injury in Mouse Model of Diabetic Kidney Disease via Nrf2/HO-1/NQO1 Signaling Pathway
Yifei ZHANG ; Xuehui BAI ; Zijing CAO ; Zeyu ZHANG ; Jingyi TANG ; Junyu XI ; Shujiao ZHANG ; Shuaixing ZHANG ; Yiran XIE ; Yuqi WU ; Zhongjie LIU ; Weijing LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):41-51
ObjectiveTo investigate the effect and mechanism of Yishen Tongluo prescription in protecting mice from oxidative stress injury in diabetic kidney disease (DKD) via the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/NAD(P)H quinone oxidoreductase 1 (NQO1) signaling pathway. MethodsSpecific pathogen-free (SPF) male C57BL/6 mice were assigned into a control group (n=10) and a modeling group (n=50). The DKD model was established by intraperitoneal injection of streptozotocin. The mice in the modeling group were randomized into a model group, a semaglutide (40 μg·kg-1) group, and high-, medium-, and low-dose (18.2, 9.1, 4.55 g·kg-1, respectively) Yishen Tongluo prescription groups, with 10 mice in each group. The treatment lasted for 12 weeks. Blood glucose and 24-h urine protein levels were measured, and the kidney index (KI) was calculated. Serum levels of creatinine (SCr), blood urea nitrogen (BUN), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were assessed. The pathological changes in the renal tissue were evaluated by hematoxylin-eosin, periodic acid-Schiff, periodic acid-silver methenamine, and Masson’s trichrome staining. Enzyme-linked immunosorbent assay kits were used to measure the levels of β2-microglobulin (β2-MG), neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), liver fatty acid-binding protein (L-FABP), nitric oxide synthase (NOS), glutathione (GSH), total antioxidant capacity (T-AOC), and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Immunohistochemical staining was performed to examine the expression of Kelch-like ECH-associated protein 1 (Keap1) and malondialdehyde (MDA). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of factors in the Nrf2/HO-1/NQO1 signaling pathway. ResultsCompared with the control group, the DKD model group showed rises in blood glucose, 24-h urine protein, KI, SCr, BUN, and ALT levels, along with glomerular hypertrophy, renal tubular dilation, thickened basement membrane, mesangial expansion, and collagen deposition. Additionally, the model group showed elevated levels of β2-MG, NGAL, KIM-1, L-FABP, NOS, and 8-OHdG, lowered levels of GSH and T-AOC, up-regulated expression of MDA and Keap1, and down-regulated expression of Nrf2, HO-1, NQO1, and glutamate-cysteine ligase catalytic subunit (GCLC) (P<0.05). Compared with the model group, the semaglutide group and the medium- and high-dose Yishen Tongluo prescription groups showed reductions in blood glucose, 24-h urine protein, KI, SCr, BUN, and ALT levels, along with alleviated pathological injuries in the renal tissue. In addition, the three groups showed lowered levels of β2-MG, NGAL, KIM-1, L-FABP, NOS, and 8-OHdG, elevated levels of GSH and T-AOC, down-regulated expression of MDA and Keap1, and up-regulated expression of Nrf2, HO-1, NQO1, and GCLC (P<0.05). ConclusionYishen Tongluo prescription exerts renoprotective effects in the mouse model of DKD by modulating the Nrf2/HO-1/NQO1 signaling pathway, mitigating oxidative stress, and reducing renal tubular injuries.
3.Simultaneous determination of 22 volatile organic compounds in urine of automotive spray painting workers using static headspace gas chromatography-mass spectrometry
Mengmeng LIU ; Junyu PAN ; Baofeng LIU ; Qiang ZENG
Journal of Environmental and Occupational Medicine 2025;42(5):608-615
Background Automotive repair workers are exposed to a wide variety of volatile organic compounds (VOCs) during painting operations, which poses significant health risks. Biomonitoring can directly reflect the internal body burden of these compounds. Therefore, it is essential to develop an analytical method for simultaneous determination of VOCs in the urine of automotive spray painting workers. Objective To establish a static headspace gas chromatography-mass spectrometry (GC-MS) method for simultaneously determining 22 VOCs in the urine of spray painting workers in automotive repair enterprises. Methods An automatic headspace sampler was used for the pretreatment of urine samples. The headspace conditions as well as the chromatographic and mass spectrometric conditions of static headspace GC-MS were optimized. The separation of 22 VOCs was achieved by optimizing the temperature program. Sensitivity was enhanced by optimizing the quantitative ions. The signal response of VOCs was improved by optimizing the headspace equilibrium temperature, equilibrium time, and the amount of inorganic salt added. The method's detection limit, lower limit of quantification, accuracy, precision, and stability were tested using blank urine samples spiked with standards. Additionally, the method was applied to examine 40 urine samples collected from painting workers in automotive repair enterprises in Tianjin. Results In this study, the headspace equilibrium temperature was set at 80 ℃, the equilibrium time was 30 min, and the salt addition amount was 2.0 g. A DB-624ms chromatographic column was selected for the separation of 22 VOCs. The initial temperature of heating program was 50 ℃, maintained for 15 min, and then increased to 85 ℃ at a heating rate of 10 ℃·min−1 for 10 min, followed by increasing to 90 ℃ at a heating rate of 5 ℃·min−1 for 20 min. The single ion monitoring (SIM) mode was chosen for the quantitative analysis of the 22 VOCs. The method demonstrated good linearity for determining 22 target analytes in the urine of spray painting workers, with correlation coefficients all above 0.990. The lower limits of quantification (LOQ) for the components ranged from 0.4 to 3.8 μg·L−1. The spiked recovery rates of the samples were between 80.1% and 112.1%, and the relative standard deviations (RSD) ranged from 1.5% to 9.7%. All target analytes could be stored at 4 ℃ for 5 d and at −20 ℃ for 7 d. This method was applied to evaluate urine samples from 40 spray painting workers in automotive repair enterprises in Tianjin. The positive rate of butyl acetate was 37.5%. The positive rate of xylene was 32.5%. The positive rate of toluene was 30.0%. The positive rate of isopropanol was 25.0%. The concentration range of the detected substances was from
4.Direct and rapid screening of four organic compound metabolites in urine by paper spray mass spectrometry
Junyu PAN ; Mengmeng LIU ; Qiang ZENG
Journal of Environmental and Occupational Medicine 2025;42(7):846-853
Background Currently, incidents of organic solvent poisoning are occurring frequently. Rapidly and accurately qualitative and quantitative analysis of toxic substances is crucial for the treatment of affected individuals. In recent years, many biomarker assays with good specificity and high sensitivity have been developed for the detection of exposure to organic solvents, but they cannot meet the demand for real-time and fast detection. Objective To establish a paper spray mass spectrometry method for direct and rapid detection of four organic compound metabolites (toluene diamine, 2,5-hexanedione, hippuric acid, and methylhippuric acid) in the urine of occupational populations. Methods Toluene diamine and 2,5-hexanedione were analyzed using positive ion mode, while hippuric acid and methylhippuric acid were analyzed using negative ion mode. The ion transfer tube temperature was 275 °C. The ion transfer tube voltage was 35 V. For positive ion mode, the scan range was 50-150 m/z. For negative ion mode, the scan range was 150-250 m/z. The distance from the paper substrate tip to the mass spectrometry inlet was 8 mm. The applied voltage was 3.5 kV. The spray solvent was methanol/water (90:10). The spray solvent volume was 15 μL. Under the optimized experimental conditions, both external standard and internal standard methods were used for quantitative analysis. Limit of detection, limit of quantification, accuracy, and precision of the proposed method were determined by spiking blank urine samples. To evaluate the feasibility of the method, the established approach was compared with current national standard detection methods or methods described in the literature. All methods were used to analyze 40 urine samples collected from occupationally exposed individuals (20 exposed to n-hexane and 20 exposed to toluene and xylene). Results The four biomarkers showed good linearity within their respective measurement ranges and the correlation coefficients were higher than
5.Analysis of the global competitive landscape in artificial intelligence medical device research.
Juan CHEN ; Lizi PAN ; Junyu LONG ; Nan YANG ; Fei LIU ; Yan LU ; Zhaolian OUYANG
Journal of Biomedical Engineering 2025;42(3):496-503
The objective of this study is to map the global scientific competitive landscape in the field of artificial intelligence (AI) medical devices using scientific data. A bibliometric analysis was conducted using the Web of Science Core Collection to examine global research trends in AI-based medical devices. As of the end of 2023, a total of 55 147 relevant publications were identified worldwide, with 76.6% published between 2018 and 2024. Research in this field has primarily focused on AI-assisted medical image and physiological signal analysis. At the national level, China (17 991 publications) and the United States (14 032 publications) lead in output. China has shown a rapid increase in publication volume, with its 2023 output exceeding twice that of the U.S.; however, the U.S. maintains a higher average citation per paper (China: 16.29; U.S.: 35.99). At the institutional level, seven Chinese institutions and three U.S. institutions rank among the global top ten in terms of publication volume. At the researcher level, prominent contributors include Acharya U Rajendra, Rueckert Daniel and Tian Jie, who have extensively explored AI-assisted medical imaging. Some researchers have specialized in specific imaging applications, such as Yang Xiaofeng (AI-assisted precision radiotherapy for tumors) and Shen Dinggang (brain imaging analysis). Others, including Gao Xiaorong and Ming Dong, focus on AI-assisted physiological signal analysis. The results confirm the rapid global development of AI in the medical device field, with "AI + imaging" emerging as the most mature direction. China and the U.S. maintain absolute leadership in this area-China slightly leads in publication volume, while the U.S., having started earlier, demonstrates higher research quality. Both countries host a large number of active research teams in this domain.
Artificial Intelligence
;
Bibliometrics
;
Humans
;
China
;
Equipment and Supplies
;
United States
;
Biomedical Research
6.Mendelian randomization studies on cardiometabolic factors and intracranial aneurysms: A systematic literature analysis.
Yuge WANG ; Junyu LIU ; Fang CAO ; Yuxin GUO ; Junxia YAN
Journal of Central South University(Medical Sciences) 2025;50(5):757-765
OBJECTIVES:
Intracranial aneurysm (IA) has an insidious onset, and once ruptured, it carries high rates of mortality and disability. Cardiometabolic factors may be associated with the formation and rupture of IA. This study aims to summarize the application of Mendelian randomization (MR) methods in research on cardiometabolic factors and IA, providing insights for further elucidation of IA etiology and pathogenesis.
METHODS:
Literature about MR-based IA studies published up to February 21, 2024, was retrieved from PubMed, Embase, Web of Science, CNKI, and Wanfang. Two researchers independently performed literature screening, data extraction, and quality assessment. A narrative synthesis approach was used to conduct a qualitative systematic review of the included studies.
RESULTS:
A total of 11 MR-based studies on IA published between 2017 to 2024 were included, of which 4 were rated as high quality. These studies investigated the associations between blood pressure, blood lipids, blood glucose, obesity-related indicators, and inflammatory cytokines with IA and its subtypes, though issues of duplication were noted. Four MR studies based on the same European population but using different instrumental variable selection criteria, as well as another MR study in a different European cohort, consistently identified blood pressure as a risk factor for IA and its subtypes. Findings for blood lipids, blood glucose, obesity-related indicators, and inflammatory cytokines were inconsistent across MR studies.
CONCLUSIONS
Blood pressure appears to increase the risk of IA and its subtypes. Associations between other cardiometabolic factors and IA/subtypes require further in-depth investigation. Given the inherent limitations of MR studies, causal inferences should be made cautiously in combination with other lines of evidence.
Humans
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Mendelian Randomization Analysis
;
Intracranial Aneurysm/etiology*
;
Risk Factors
;
Blood Pressure
;
Blood Glucose
;
Obesity/complications*
;
Cardiometabolic Risk Factors
;
Lipids/blood*
7.Application of domestic jingfeng single-port robotic-assisted surgery in multisite diseases of oropharyngeal-head and neck region: case series.
Ping HAN ; Faya LIANG ; Peiliang LIN ; Junyu CHEN ; Jingyi WANG ; Yanke LIU ; Xiaoming HUANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(11):1048-1056
Objective:To evaluate the safety, feasibility, and technical advantages of the domestic Jingfeng single-port robotic surgical system in transoral and transaxillary approach surgeries for multisite diseases. Methods:A retrospective analysis was conducted on the clinical data of 6 patients who underwent Jingfeng SP1000 single-port robotic-assisted surgery at our hospital from June 2025 to July 2025. They were divided into the transoral robotic surgery group(4cases) and the transaxillary approach thyroid cancer radical resection group(2cases) based on surgical approaches. The transoral robotic surgery group included extended resection of right tonsillar cancer with cervical lymph node dissection, epiglottic cyst resection, extended resection of right pyriform sinus cancer with cervical lymph node dissection, and surgery for epiglottic cyst and obstructive sleep apnea(OSA). For each case, parameters including operative time, intraoperative blood loss, perioperative complications, robotic operation performance, and postoperative recovery were recorded. Results:All 6 surgeries were successfully completed without conversion to open surgery or system failure. In the transoral robotic surgery group, the mean robotic operation time was(60.00±34.88) minutes, and the mean intraoperative blood loss was 20.00(5.75,20.00)mL. In the transaxillary robotic surgery group, the robotic operation time was respectively 60.00 and 40.00 minutes, and the intraoperative blood loss was 10.00 and 5.00 mL, respectively. One case of minor perioperative complication(in the flap dissection area) occurred, with no severe complications reported. All patients recovered smoothly after surgery, with a median follow-up of 1.9 months showing no residual lesions, recurrence, or functional impairment. Conclusion:This case series confirms the safety and feasibility of the domestic Jingfeng single-port robot in transoral and axillary approach surgeries in oropharyngeal-head and neck surgery. Its single-port design reduces trauma and the risk of robotic arm collision, adapts to minimally invasive needs, and its domestic production attribute lowers costs to facilitate popularization, providing a new option for such patients.
Humans
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Robotic Surgical Procedures/methods*
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Retrospective Studies
;
Operative Time
;
Middle Aged
;
Male
;
Female
;
Neck/surgery*
;
Sleep Apnea, Obstructive/surgery*
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Adult
;
Head and Neck Neoplasms/surgery*
;
Oropharynx/surgery*
;
Oropharyngeal Neoplasms/surgery*
8.Histopathological Insights into Demyelination and Remyelination After Spinal Cord Injury in Non-human Primates.
Junhao LIU ; Zucheng HUANG ; Kinon CHEN ; Rong LI ; Zhiping HUANG ; Junyu LIN ; Hui JIANG ; Jie LIU ; Qingan ZHU
Neuroscience Bulletin 2025;41(8):1429-1447
Demyelination and remyelination play key roles in spinal cord injury (SCI), affecting the recovery of motor and sensory functions. Research in rodent models is extensive, but the study of these processes in non-human primates is limited. Therefore, our goal was to thoroughly study the histological features of demyelination and remyelination after contusion injury of the cervical spinal cord in Macaca fascicularis. In a previous study, we created an SCI model in M. fascicularis by controlling the contusion displacement. We used Eriochrome Cyanine staining, immunohistochemical analysis, and toluidine blue staining to evaluate demyelination and remyelination. The results showed demyelination ipsilateral to the injury epicenter both rostrally and caudally, the former mainly impacting sensory pathways, while the latter primarily affected motor pathways. Toluidine blue staining showed myelin loss and axonal distension at the injury site. Schwann cell-derived myelin sheaths were only found at the center, while thinner myelin sheaths from oligodendrocytes were seen at the center and surrounding areas. Our study showed that long-lasting demyelination occurs in the spinal cord of M. fascicularis after SCI, with oligodendrocytes and Schwann cells playing a significant role in myelin sheath formation at the injury site.
Animals
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Spinal Cord Injuries/physiopathology*
;
Demyelinating Diseases/etiology*
;
Remyelination/physiology*
;
Macaca fascicularis
;
Disease Models, Animal
;
Myelin Sheath/pathology*
;
Oligodendroglia/pathology*
;
Schwann Cells/pathology*
;
Female
;
Spinal Cord/pathology*
;
Axons/pathology*
9.Discussion on the application status and method improvement of gas chromatography in occupational health inspection standards in China
Junyu PAN ; Mengmeng LIU ; Qiang ZENG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(3):213-217
As a rapid, accurate and efficient analytical technique, gas chromatography is widely used in the detection of volatile organic compounds and inorganic small molecule toxins, and it is the main analytical method in the national testing standards for occupational health. The existing effective national standards of gas chromatography for the detection of some substances have low column efficiency, high toxicity of reagents, poor correlation of the standard curve and low desorption efficiency and other problems, some of which can be solved through method improvement. At the same time, with the use of new materials and new processes, new types of toxic substances are emerging, and there are still many occupational disease hazards of limited value without supporting detection methods, gas chromatography can be applied to the detection of some toxic substances to better complement the vacancy of China's occupational health detection methods. This paper analyzes the current situation of the application of gas chromatography in occupational health testing standards, discusses the improvement of some of these methods, and helps to promote the application and development of gas chromatography in occupational health testing.
10.Visualization analysis of literature on sudden poisoning incidents in China from 2013 to 2022 based on CiteSpace and VOSviewer
Mengmeng LIU ; Junyu PAN ; Qiang ZENG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(11):831-837
Objective:To use CiteSpace and VOSviewer software to perform visualization analysis on research literature on sudden poisoning incidents in China from 2013 to 2022, in order to grasp the research hotspots and priorities, current research development status and trend of sudden poisoning incidents, and provide reference and ideas for subsequent research.Methods:In October 2023, literature related to sudden poisoning incidents was searched using the China National Knowledge Infrastructure and Wanfang databases. The bibliometric analysis software CiteSpace 6.1.R2 and VOSviewer 1.6.19 were used to visually analyze factors such as authors, institutions, journals, regions, and keywords in the relevant literature.Results:A total of 2578 valid literatures were included, and the overall number of published literatures showed a discontinuous downward trend in recent 10 years. The relevant literature was distributed in 310 journals. Among the top 10 institutions with the highest number of publications, 8 were centers for disease control and prevention at all levels, and the institution with the highest number of publications was the Chinese Center for Disease Control and Prevention (208 articles), but the cooperation between institutions was not obvious. The 2578 articles involved a total of 6549 authors (a total of 8572 appearances, with an average of 1.31 times per person), with a collaboration rate of 2.54 (6549/2578) and 2153 co-authored articles (with 2 or more authors in the literature), with a co-authorship rate of 83.51%. 643 articles (24.94%) received funding support. The region with the highest number of publications was Guangdong Province, with a total of 380 articles (14.74%). Keyword highlighting analysis showed that the research heat of pesticide poisoning, toxic mushrooms, disease characteristics, outbreak events, traceability analysis, outbreaks, and occupational diseases continued to 2022.Conclusion:There are many literatures on sudden poisoning incidents in China, so it is necessary to strengthen the cooperation and communication between institutions and authors. In practical work, the ability of prevention, control, and detection of poisons should be continuously improved to reduce the incidence and degree of harm caused by sudden poisoning incidents.


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