In the present study, a mouse model of coronary artery ligation was employed to evaluate the effects of Jiming Powder on mitophagy in the mouse model of myocardial infarction and elucidate its underlying mechanisms. A mouse model of myocardial infarction post heart failure was constructed by ligating the left anterior descending branch of the coronary artery. The therapeutic efficacy of Jiming Powder was assessed from multiple perspectives, including ultrasonographic imaging, hematoxylin-eosin(HE) staining, Masson staining, and serum cardiac enzyme profiling. Dihydroethidium(DHE) staining was employed to evaluate the oxidative stress levels in the hearts of mice from each group. Mitophagy levels were assessed by scanning electron microscopy and immunofluorescence co-localization. Western blot was employed to determine the levels of key proteins involved in mitophagy, including Bcl-2-interacting protein beclin 1(BECN1), sequestosome 1(SQSTM1), microtubule-associated protein 1 light chain 3 beta(LC3B), PTEN-induced putative kinase 1(PINK1), phospho-Parkinson disease protein(p-Parkin), and Parkinson disease protein(Parkin). The results demonstrated that compared with the model group, high and low doses of Jiming Powder significantly reduced the left ventricular internal diameter in systole(LVIDs) and left ventricular internal diameter in diastole(LVIDd) and markedly improved the left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS), effectively improving the cardiac function in post-myocardial infarction mice. Jiming Powder effectively reduced the levels of myocardial injury markers such as creatine kinase(CK), creatine kinase isoenzyme(CK-MB), and lactate dehydrogenase(LDH), thereby protecting ischemic myocardium. HE staining revealed that Jiming Powder attenuated inflammatory cell infiltration after myocardial infarction. Masson staining indicated that Jiming Powder effectively inhibited ventricular remodeling. Western blot results showed that Jiming Powder activated the PINK1-Parkin pathway, up-regulated the protein level of BECN1, down-regulated the protein level of SQSTM1, and increased the LC3Ⅱ/LC3Ⅰ ratio to promote mitophagy. In conclusion, Jiming Powder exerts therapeutic effects on myocardial infarction by inhibiting ventricular remodeling. The findings pave the way for subsequent pharmacological studies on the active components of Jiming Powder.
Animals ; Myocardial Infarction/physiopathology* ; Mitophagy/drug effects* ; Mice ; Drugs, Chinese Herbal/administration & dosage* ; Protein Kinases/genetics* ; Male ; Ubiquitin-Protein Ligases/genetics* ; Humans ; Disease Models, Animal ; Mice, Inbred C57BL ; Signal Transduction/drug effects*

This study employed a mouse model of coronary artery ligation to assess the effect and mechanism of Jiming Powder on mitochondrial autophagy in mice with myocardial infarction. The mouse model of heart failure post-myocardial infarction was established by ligating the left anterior descending coronary artery. The pharmacological efficacy of Jiming Powder was evaluated through echocardiographic imaging, hematoxylin-eosin(HE) staining, and Masson staining. The levels of malondialdehyde(MDA), Fe~(2+), reduced glutathione(GSH), and superoxide dismutase(SOD) in heart tissues, as well as MDA immunofluorescence of heart tissues, were measured to assess lipid peroxidation and Fe~(2+) levels in the hearts of mice in different groups. Ferroptosis levels in the groups were evaluated using scanning electron microscopy and Prussian blue staining. Western blot analysis was conducted to detect the levels of key ferroptosis-related proteins, including nuclear factor erythroid 2-related factor 2(NRF2), ferritin heavy chain(FTH), glutathione peroxidase 4(GPX4), solute carrier family 7 member 11(SLC7A11), heme oxygenase 1(HO-1), and Kelch-like ECH-associated protein 1(KEAP1). The results showed that compared with the model group, both the high-and low-dose Jiming Powder groups exhibited significantly reduced left ventricular internal diameter in systole(LVIDs) and left ventricular internal diameter in diastole(LVIDd), while the left ventricular ejection fraction(EF) and left ventricular fractional shortening(FS) were significantly improved, effectively enhancing cardiac function in mice post-myocardial infarction. HE staining revealed that Jiming Powder attenuated myocardial inflammatory cell infiltration post-infarction, and Masson staining indicated that Jiming Powder effectively reduced fibrosis in the infarct margin area. Treatment with Jiming Powder reduced the levels of MDA and Fe~(2+), indicators of lipid peroxidation post-myocardial infarction, while increasing GSH and SOD levels, thus protecting ischemic myocardium. Western blot results demonstrated that Jiming Powder reduced KEAP1 protein accumulation, activated the NRF2/HO-1/GPX4 pathway, and up-regulated the protein expression of FTH and SLC7A11, exerting an inhibitory effect on ferroptosis. This study reveals that Jiming Powder exerts a therapeutic effect on myocardial infarction by inhibiting ferroptosis through the NRF2/HO-1/GPX4 pathway, providing a foundation for subsequent research on the pharmacological effects of Jiming Powder.
Animals ; Ferroptosis/drug effects* ; Myocardial Infarction/physiopathology* ; NF-E2-Related Factor 2/genetics* ; Mice ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Heme Oxygenase-1/genetics* ; Phospholipid Hydroperoxide Glutathione Peroxidase/genetics* ; Humans ; Mice, Inbred C57BL ; Signal Transduction/drug effects* ; Disease Models, Animal

Neuropathic pain, often featuring allodynia, imposes significant physical and psychological burdens on patients, with limited treatments due to unclear central mechanisms. Addressing this challenge remains a crucial unsolved issue in pain medicine. Our previous study, using protein kinase C gamma (PKCγ)-tdTomato mice, highlights the spinal feedforward inhibitory circuit involving PKCγ neurons in gating neuropathic allodynia. However, the regulatory mechanisms governing this circuit necessitate further elucidation. We used diverse transgenic mice and advanced techniques to uncover the regulatory role of the descending serotonin (5-HT) facilitation system on spinal PKCγ neurons. Our findings revealed that 5-HT neurons from the rostral ventromedial medulla hyperpolarize spinal inhibitory interneurons via 5-HT_2C receptors, disinhibiting the feedforward inhibitory circuit involving PKCγ neurons and exacerbating allodynia. Inhibiting spinal 5-HT_2C receptors restored the feedforward inhibitory circuit, effectively preventing neuropathic allodynia. These insights offer promising therapeutic targets for neuropathic allodynia management, emphasizing the potential of spinal 5-HT_2C receptors as a novel avenue for intervention.
Animals ; Neuralgia/physiopathology* ; Protein Kinase C/metabolism* ; Receptor, Serotonin, 5-HT2C/metabolism* ; Hyperalgesia/physiopathology* ; Mice, Transgenic ; Mice ; Spinal Cord/metabolism* ; Serotonin/metabolism* ; Male ; Neurons/metabolism* ; Mice, Inbred C57BL

Objective:This study aimed to evaluate to evaluate the performance of equilibrium dialysis combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the accurate measurement of free testosterone in clinical samples. Mthods We conducted a prospective observational study using 161 serum samples from healthy women of reproductive 26(24, 32)years at the Gynecology Outpatient department of Peking Union Medical College Hospital from June to September 2024, and their concentrations were determined. In this study, after equilibrium dialysis of serum samples, free testosterone was extracted from the dialysate using a magnetic bead-based method. It was then directly derivatized using hydroxylamine hydrochloride in situ after elution from the magnetic bead and further quantified by LC-MS/MS.Method:validation assessed linearity, limit of quantification (LOQ), precision, accuracy, matrix effects, and carryover according to established guidelines. Data were analyzed using Origin 2019 and WPS Office 2019.Results:The method demonstrated excellent linearity ( R2>0.99) across 1-250 pg/ml with an LOQ of 1 pg/ml. The coefficients of variation for both intra-day and inter-day imprecision were less than 10% while recovery rates ranged from 92.60% to 99.10%. Matrix effect deviations were all within the range of 6% and carryover was negligible. Conclusions:In this study, the established method of magnetic bead-based extraction followed by in situ derivatization combined with liquid chromatography-tandem mass spectrometry performed well, and could be further applied to the detection of free testosterone concentration in childbearing age women.

A highly efficient oxidase-mimetic silver phosphate/nickel hydroxystannate(Ag3PO4/NiSn(OH)6)composite was synthesized via a precipitation method using nickel hydroxystannate(NiSn(OH)6)as the support.The abundant hydroxyl groups(—OH)on NiSn(OH)6 not only provided nucleation sites for Ag3PO4 nanoparticles but also improved their dispersion and overall material stability.Based on oxidase-like activity of Ag3PO4/NiSn(OH)6 and inhibitory effect of sodium dodecylbenzenesulfonate(SDBS)on this catalytic activity,a novel colorimetric sensing method for SDBS detection was developed.Under optimized experimental conditions,the method exhibited a linear range of 3.69-42.7 μmol/L,with a detection limit of 0.135 μmol/L(S/N=3).The regression equation was ΔA652=0.01125C(μmol/L)+0.1498,with a correlation coefficient(R2)of 0.992.Practical application in dishwashing liquid analysis achieved satisfactory recoveries of 96.9%-106.4%,demonstrating the method's reliability for real sample detection.

In the 1950s and 1960s,Japan's implementation of policies prioritizing economic develop-ment caused a lack of effective supervision over the discharge of industrial wastewater and exhaust gases,which led to the occurrence of the"Four Major Pollution Diseases",including Minamata disease,causing serious social and public health problems.To more effectively address public nuisances and pro-vide compensation to victims,the Japanese government gradually established an environmental damage compensation system with administrative relief characteristics since the 1970s.Through long-term prac-tice and system optimization,this system has evolved into a mature institutional framework with a clear division of labor and efficient collaboration.This paper systematically reviews the development process of Japan's environmental damage compensation system and deeply analyzes its legal frame-work and supporting policies,aiming to provide useful references for the construction and improve-ment of China's environmental damage compensation system.Meanwhile,through the case analysis of Minamata disease,the paper explores the specific mechanisms and effects in the compensation practices,further revealing the system's operational characteristics and implications,and providing a reference ba-sis for the construction of China's environmental governance legal system.

Objective To investigate the genetic differences among different populations based on 13 autosomal STR loci in CODIS core.Methods Data of 13 autosomal STR loci(CSF1PO,FGA,THO1,TPOX,vWA,D3S1358,D5S818,D7S820,D8S1179,D13S317,D16S539,D18S51,D21S11)were collected from 95 populations in scientific journals between 1999 and 2021,soursed from the PubMed database,which had been published.Allele frequencies of loci were sorted out and forensic genetic parameters including gene differentiation coefficient(Gst),total heterozygosity(Ht),subpopula-tion heterozygosity(Hs)values,and Nei's DA genetic distance were calculated.Principal component analysis,phylogenetic tree,and multidimensional scale analysis were conducted to assess population ge-netic structure.Results A total of 265 alleles were detected at the 13 STR loci in these 95 popula-tions.The mean values of Gst,Ht,and Hs were 0.023 247,0.797 915 and 0.779 365.Population genetic analyses reflected significant differences among populations from Asia,Africa and Europe.In Asian populations,there was a certain degree of distinction between mainland and island populations;the Han population showed a certain degree of distinction with surrounding populations in mainland;while within the Han population,there were two distinct clusters formed by the northern Han and the south-ern Han.Conclusion The 13 autosomal STR loci in CODIS core demonstrate potential value for popu-lation identification across different groups,and may be used for the differentiation of ethnic groups,among different continental populations.

Objective:To investigate genetic findings and pregnancy outcomes in fetuses with omphalocele.Methods:This retrospective study analyzed data from 502 fetuses with prenatally diagnosed omphalocele who underwent genetic testing at Guangzhou Women and Children's Medical Center and Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region between January 2014 and March 2024. Testing methods included karyotyping, chromosomal microarray analysis (CMA), methylation-specific multiplex ligation-dependent probe amplification, and whole-exome sequencing (WES). Cases were categorized as non-isolated ( n=340) or isolated ( n=162) based on ultrasound findings. Differences in genetic abnormality detection rates and pregnancy outcomes were analyzed using Mann-Whitney U test, independent samples t-test, and Chi-square test (or Fisher's exact test). Results:Among 502 fetuses, karyotyping plus CMA detected chromosomal abnormalities in 223 cases (44.4%, 223/502), including trisomy 18 (57.0%, 127/223) and trisomy 13 (23.3%, 52/223). CMA additionally identified nine pathogenic copy number variations (1.8%, 9/502) and five uniparental disomies (1.0%, 5/502), increasing the total diagnostic yield from 44.4% to 47.2%. The genetic abnormality rate was significantly lower in isolated (14.8%, 24/162) versus non-isolated omphalocele (64.7%, 220/340) ( χ2=109.34, P<0.001). WES detected variants in nine of 16 karyotype/CMA-negative cases, including five pathogenic variants involving PIK3CA and CDKN1C. Eight imprinting disorders (1.6%, 8/502) were identified, including five Beckwith-Wiedemann syndrome cases. Among 499 cases with follow-up, 401 (80.4%, 401/499) underwent pregnancy termination. Live birth rate was higher in isolated versus non-isolated groups [42.5% (69/162) vs. 8.5% (29/340), χ2=77.67, P<0.001]. Three cases were lost to follow-up. The one-year survival rate was 93.9% (92/98) in live-born infants. Conclusion:Aneuploidy (particularly trisomy 18) is the primary genetic etiology of omphalocele. CMA and WES significantly improve diagnostic yield. Isolated omphalocele has a more favorable prognosis, while non-isolated cases show significantly higher genetic abnormality rates. A stratified testing strategy is recommended: karyotyping plus CMA for isolated cases and prioritization of WES for multiple anomalies.

Objective:To analyze the correlation between blood glucose index and peripheral neuropathy in type 2 diabetes patients under continuous blood glucose monitoring.Methods:A total of 110 patients with type 2 diabetes admitted to the Lu′an Hospital Affiliated to Anhui Medical University from November 2022 to January 2024 were retrospectively observed, and the blood glucose indexes of patients were monitored by continuous glucose meter [glucose time in range (TIR), mean blood glucose (MBG), estimated hemoglobin A 1c (eHbA 1c), blood glucose standard deviation (SD), coefficient of variation (CV), largest amplitude of glycemic excursions (LAGE), mean amplitude of glycemic excursions (MAGE), means of daily differences (MODD), etc ]. All patients with type 2 diabetes were grouped according to whether they had peripheral neuropathy or not, and were divided into the developing group and the non-developing group. General data and blood glucose indexes of the two groups were collected and sorted out. Receiver operating characteristic (ROC) curve was used to analyze the value of various blood glucose indicators in predicting peripheral neuropathy in patients with type 2 diabetes. Spearman correlation analysis was also used to evaluate the correlation between blood glucose index and peripheral neuropathy. Results:The disease course, TIR, MBG, eHbA 1c, SD, CV, LAGE, MAGE and MODD of the occurrence group were significantly different from those of the non-occurrence group (all P<0.05). By plotting the ROC curve, It was found that the area under the curve (AUC) of TIR, MBG, eHbA 1c, SD, CV, LAGE, MAGE and MODD predicting peripheral neuropathy in patients with type 2 diabetes were 0.942, 0.840, 0.705, 0.759, 0.819, 0.813, 0.857 and 0.677, respectively. The AUC of the combined prediction was 0.971(95% CI: 0.946-0.997), which was higher than that of the single indicator (all P<0.05). Spearman correlation assessment showed a negative correlation between TIR and peripheral neuropathy ( r=-0.738, P<0.05). MBG, eHbA 1c, SD, CV, LAGE, MAGE and MODD were positively correlated with peripheral neuropathy ( r=0.554, 0.376, 0.452, 0.490, 0.527, 0.625, 0.272, all P<0.05). Conclusions:Based on continuous blood glucose monitoring, the blood glucose index of type 2 diabetes patients is closely related to peripheral neuropathy, and the above blood glucose index can accurately predict peripheral neuropathy, providing a reference for reducing or preventing the occurrence of peripheral neuropathy.

Objective:To analyze the correlation between blood glucose index and peripheral neuropathy in type 2 diabetes patients under continuous blood glucose monitoring.Methods:A total of 110 patients with type 2 diabetes admitted to the Lu′an Hospital Affiliated to Anhui Medical University from November 2022 to January 2024 were retrospectively observed, and the blood glucose indexes of patients were monitored by continuous glucose meter [glucose time in range (TIR), mean blood glucose (MBG), estimated hemoglobin A 1c (eHbA 1c), blood glucose standard deviation (SD), coefficient of variation (CV), largest amplitude of glycemic excursions (LAGE), mean amplitude of glycemic excursions (MAGE), means of daily differences (MODD), etc ]. All patients with type 2 diabetes were grouped according to whether they had peripheral neuropathy or not, and were divided into the developing group and the non-developing group. General data and blood glucose indexes of the two groups were collected and sorted out. Receiver operating characteristic (ROC) curve was used to analyze the value of various blood glucose indicators in predicting peripheral neuropathy in patients with type 2 diabetes. Spearman correlation analysis was also used to evaluate the correlation between blood glucose index and peripheral neuropathy. Results:The disease course, TIR, MBG, eHbA 1c, SD, CV, LAGE, MAGE and MODD of the occurrence group were significantly different from those of the non-occurrence group (all P<0.05). By plotting the ROC curve, It was found that the area under the curve (AUC) of TIR, MBG, eHbA 1c, SD, CV, LAGE, MAGE and MODD predicting peripheral neuropathy in patients with type 2 diabetes were 0.942, 0.840, 0.705, 0.759, 0.819, 0.813, 0.857 and 0.677, respectively. The AUC of the combined prediction was 0.971(95% CI: 0.946-0.997), which was higher than that of the single indicator (all P<0.05). Spearman correlation assessment showed a negative correlation between TIR and peripheral neuropathy ( r=-0.738, P<0.05). MBG, eHbA 1c, SD, CV, LAGE, MAGE and MODD were positively correlated with peripheral neuropathy ( r=0.554, 0.376, 0.452, 0.490, 0.527, 0.625, 0.272, all P<0.05). Conclusions:Based on continuous blood glucose monitoring, the blood glucose index of type 2 diabetes patients is closely related to peripheral neuropathy, and the above blood glucose index can accurately predict peripheral neuropathy, providing a reference for reducing or preventing the occurrence of peripheral neuropathy.