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.

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.

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 −1. Conclusion The established simultaneous determination protocol of 22 VOCs by static headspace- GC-MS is characterized by its simplicity, high sensitivity, excellent selectivity, and high accuracy. It is suitable for urine samples of spray painting workers in automotive repair industry.

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 0.9990. The limits of detection of the method were in the range of 0.00027 to 0.020 μg·mL⁻¹, and the limits of quantification were in the range of 0.0009 to 0.067 μg·mL⁻¹. The spiked recoveries at low, medium, and high concentrations ranged from 96.5% to 106.9%, with relative standard deviations (RSDs) between 5.6% and 9.7%. When applying the method to 40 urine samples from occupationally exposed individuals, the detection rates for toluene diamine, 2,5-hexanedione, hippuric acid, and methylhippuric acid were 0%, 45%, 20%, and 37.5%, respectively. For biomarkers detected by both this method and the reference methods, the detection values were generally consistent. The analysis time per sample was approximately 1-2 min using paper spray mass spectrometry, compared to 1-2 h for reference methods. Conclusion The paper spray mass spectrometry method established in this study for detecting four organic compound metabolites in urine is characterized by its simplicity, rapidity, efficiency, and high accuracy. It can obtain results within 1-2 min, which significantly improves the timeliness of detection. It is suitable for direct and rapid screening of four organic compound metabolites in urine, and can provide technical support for the diagnosis and treatment of organic solution poisoning.

Ring finger protein 213(RNF213)was first identified as a susceptible gene for Moyamoya disease,and also found to be associated with various vascular related diseases such as intracranial artery occlusion/stenosis,ischaemic stroke,intracranial aneurysm,cerebral arteriovenous malformation,etc.This article systematically reviewed the literature of RNF213,providing an overview of the association and possible mechanisms between RNF213 variation and related diseases,in order to provide reference for the prevention,diagnosis,and treatment of related diseases.

Objective: This study examined the gut bacterial communities of dogs from different breeds, all kept under identical domestication conditions. Methods: Noninvasive sampling and 16S rRNA high-throughput sequencing were used to compare the composition and function of the gut microbiota of three dog breeds: the Chinese Kunming dog (CKD), German Shepherd dog (GSD), and Belgian Malinois dog (BMD). Results: The gut microbiota of the three dog breeds consisted of 257 species across 146 genera, 60 families, 35 orders, 15 classes, and 10 phyla. The dominant bacterial phyla across the three breeds were Firmicutes (57.44%), Fusobacteriota (28.86%), and Bacteroidota (7.63%), while the dominant bacterial genera across the three breeds were Peptostreptococcus (21.08%), Fusobacterium (18.50%), Lactobacillus (12.37%), and Cetobacter (10.29%). Further analysis revealed significant differences in the intestinal flora of the three breeds at the phylum and genus levels. The intestinal flora of BMD was significantly richer than that of CKD and GSD. The functional prediction and Kyoto Encyclopedia of Genes and Genomes analysis showed that the primary functions of the gut microbiota in these breeds were similar, with significant enrichment in various metabolic pathways, including carbohydrate and amino acid metabolism, secondary metabolite biosynthesis, and microbial metabolism in different environments. The intestinal flora of these breeds also played a crucial role in genetic information processing, including transcription, translation, replication, and material transport. Conclusions and Relevance: These results provide novel insights into the intestinal flora of intervention dogs and suggest novel methods to improve their health status, which help increase microbial diversity and normalize metabolite production in diseased dogs.

Objective:To investigate the predictive value of differential distribution of peripheral lymphocyte subsets before and after the first 131I treatment on the therapeutic response to 131I treatment in patients with papillary thyroid cancer (PTC). Methods:A retrospective study was conducted on 46 PTC patients (16 males, 30 females, age 20-77 years) who underwent total thyroidectomy and received 131I treatment between January 2021 and August 2021 in First Hospital of Shanxi Medical University. Peripheral blood lymphocyte subsets (T, B, CD4 + T, CD8 + T, natural killer (NK), helper T (Th)1, Th2, Th17, and regulatory T (Treg) cells) were measured 1-2 d before and 30 d after 131I treatment. Based on serological and imaging evidence, therapeutic response at 6-12 months post- 131I therapy was categorized as either excellent response (ER) or non-excellent response (NER). Differences of preablative stimulated thyroglobulin (psTg) and clinical baseline characteristics between two groups were assessed by using independent-sample t test, paired t test, or Mann-Whitney U test. Predictive value of lymphocyte subsets before and after 131I treatment for therapeutic response was assessed through logistic regression analysis, ROC curve analysis, and decision curve analysis (DCA). Results:In ER group ( n=33) and NER group ( n=13), most lymphocyte subsets showed different degrees of reduction 30 d after 131I treatment compared to before 131I treatment, such as T, B, CD4 + T and Th1 cells in ER group, as well as T, B, CD4 + T, Th1, Th2, Th17, and Treg cells in NER group ( t values: 2.41-9.57, all P<0.05). Before 131I treatment, NER group had significantly higher levels of psTg, Th2, Th17, and Treg cells compared to the ER group ( t values: from -3.32 to -2.48, U=29.00, all P<0.05). After 131I treatment, most of lymphocyte subsets in NER group (T, B, CD4 + T, CD8 + T, Th1 and Treg cells) showed higher trend than those in ER group but without statistical significances ( t values: from -1.12 to -0.06, all P>0.05). Th2 cells before 131I treatment (odds ratio ( OR)=25.00, 95% CI: 1.36-459.10, P=0.030) was identified as a risk factor for NER. ROC curve analysis indicated that AUCs of psTg and Th2 cells for predicting therapeutic response were 0.932 and 0.790, respectively, which was 0.958 for the combined psTg and Th2 cells. DCA showed that within the threshold probability range of 10%-60%, the curves for psTg, Th2 cells, and the combined psTg and Th2 cells were all higher than the extreme curve, suggesting good effect. Conclusions:Most lymphocyte subsets decrease to varying degrees, and NER group shows a significant decrease 30 d after 131I treatment. Th2 cells may be a risk factor for poor response to 131I treatment, providing a certain value in predicting the therapeutic response to 131I treatment.

Objective:To examine the prevalence of frailty among elderly patients in the emergency department and to investigate the potential relationship between vitamin D nutritional status and frailty.Methods:This study collected clinical data from elderly patients aged over 65 years in the emergency intensive care unit and emergency observation ward of Beijing Bo'Ai Hospital from January to September 2021.The data included blood routine, biochemical indicators, circulating interleukin-6, cortisol, thyrotropin, and 25-hydroxyvitamin D[25(OH)D], which were detected within 24 hours after enrollment.Additionally, the Frailty Screening Questionnaire(FSQ), FRAIL scale, and Clinical Frailty Scale(CFS)were used to score the patients.Based on the scores, the patients were divided into frail or non-frail groups, and the prevalence of frailty was reported accordingly using the criteria of the aforementioned scales.The consistency of the three scales was evaluated using the Spearman rank test and Kappa coefficient.We compared the differences in clinical data and laboratory indicators of patients between the frail and non-frail groups.Additionally, we used a multivariable Logistic regression model to analyze the association between vitamin D nutritional status and frailty.We also analyzed the prevalence of frailty in different vitamin D nutritional statuses and evaluated the predictive ability of serum 25(OH)D for frailty using the receiver operating characteristic(ROC)curve.Results:A total of 317 patients were included in the study.The prevalence of frailty in elderly patients in the emergency department was found to be 47.0%, 55.2%, and 69.4% according to the FSQ, FRAIL, and CFS scales, respectively.The study evaluated the consistency of these three scales, revealing a Spearman rank correlation coefficient of 0.761(95% CI: 0.715-0.806, P<0.001)and a Kappa coefficient of 0.536(95% CI: 0.451-0.621, P<0.001)between FSQ and FRAIL, which were the highest correlations observed.Logistic regression analysis, after adjusting for age, gender, BMI, and other factors, indicated that vitamin D deficiency( OR=5.994, 95% CI: 1.232-29.169, P=0.027)was independently associated with an increased prevalence of frailty as defined by FSQ criteria.The prevalence of frailty increased with the severity of vitamin D malnutrition.In the vitamin D deficiency group, the prevalence was higher compared to the vitamin D insufficiency and sufficiency groups( P<0.05 for all).The area under the ROC curves(AUCs)of serum 25(OH)D levels to predict frailty, as defined by FSQ, FRAIL, and CFS, were 0.806(95% CI: 0.744-0.868), 0.748(95% CI: 0.679-0.817), and 0.768(95% CI: 0.701-0.826)( P<0.001 for all).The optimal cut-off values were 12.0, 9.76, and 11.65 μg/L, respectively, yielding a Youden index of 0.553, 0.419, and 0.462. Conclusions:FSQ, FRAIL, and CFS demonstrated a strong level of consistency in assessing frailty.Additionally, serum 25(OH)D can serve as an independent predictor of frailty, aiding in the identification of frail individuals and enhancing the risk stratification of elderly patients in the emergency department.

Objective: This study examined the gut bacterial communities of dogs from different breeds, all kept under identical domestication conditions. Methods: Noninvasive sampling and 16S rRNA high-throughput sequencing were used to compare the composition and function of the gut microbiota of three dog breeds: the Chinese Kunming dog (CKD), German Shepherd dog (GSD), and Belgian Malinois dog (BMD). Results: The gut microbiota of the three dog breeds consisted of 257 species across 146 genera, 60 families, 35 orders, 15 classes, and 10 phyla. The dominant bacterial phyla across the three breeds were Firmicutes (57.44%), Fusobacteriota (28.86%), and Bacteroidota (7.63%), while the dominant bacterial genera across the three breeds were Peptostreptococcus (21.08%), Fusobacterium (18.50%), Lactobacillus (12.37%), and Cetobacter (10.29%). Further analysis revealed significant differences in the intestinal flora of the three breeds at the phylum and genus levels. The intestinal flora of BMD was significantly richer than that of CKD and GSD. The functional prediction and Kyoto Encyclopedia of Genes and Genomes analysis showed that the primary functions of the gut microbiota in these breeds were similar, with significant enrichment in various metabolic pathways, including carbohydrate and amino acid metabolism, secondary metabolite biosynthesis, and microbial metabolism in different environments. The intestinal flora of these breeds also played a crucial role in genetic information processing, including transcription, translation, replication, and material transport. Conclusions and Relevance: These results provide novel insights into the intestinal flora of intervention dogs and suggest novel methods to improve their health status, which help increase microbial diversity and normalize metabolite production in diseased dogs.

Traumatic supraorbital fissure syndrome (TSOFS) is a symptom complex caused by nerve entrapment in the supraorbital fissure after skull base trauma. If the compressed cranial nerve in the supraorbital fissure is not decompressed surgically, ptosis, diplopia and eye movement disorder may exist for a long time and seriously affect the patients′ quality of life. Since its overall incidence is not high, it is not familiarized with the majority of neurosurgeons and some TSOFS may be complicated with skull base vascular injury. If the supraorbital fissure surgery is performed without treatment of vascular injury, it may cause massive hemorrhage, and disability and even life-threatening in severe cases. At present, there is no consensus or guideline on the diagnosis and treatment of TSOFS that can be referred to both domestically and internationally. To improve the understanding of TSOFS among clinical physicians and establish standardized diagnosis and treatment plans, the Skull Base Trauma Group of the Neurorepair Professional Committee of the Chinese Medical Doctor Association, Neurotrauma Group of the Neurosurgery Branch of the Chinese Medical Association, Neurotrauma Group of the Traumatology Branch of the Chinese Medical Association, and Editorial Committee of Chinese Journal of Trauma organized relevant experts to formulate Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome ( version 2024) based on evidence of evidence-based medicine and clinical experience of diagnosis and treatment. This consensus puts forward 12 recommendations on the diagnosis, classification, treatment, efficacy evaluation and follow-up of TSOFS, aiming to provide references for neurosurgeons from hospitals of all levels to standardize the diagnosis and treatment of TSOFS.