ObjectiveThis study aimed to investigate the mechanism by which Luoshi Neiyi prescription treats endometriosis （EMs） through regulating ferroptosis， and to screen key metabolites and analyze their association with ferroptosis. MethodsClinical samples of normal endometrium from patients without EMs and eutopic and ectopic endometrium from EMs patients （10 cases each） were collected and divided into control group， eutopic group， and EMs group. Hematoxylin-eosin （HE） staining was performed to observe ectopic lesions of EMs. Immunohistochemistry was used to detect the expression of solute carrier family 7 member 11 （SLC7A11） and glutathione peroxidase 4 （GPX4）. Enzyme-linked immunosorbent assay （ELISA） was adopted to determine the levels of malondialdehyde （MDA）， ferrous ion （Fe²⁺）， GPX4 and glutathione （GSH） in endometrial tissues， as well as serum levels of Fe²⁺， GPX4 and GSH. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of SLC7A11， GPX4， transferrin receptor （TFR） and ferritin heavy chain 1 （FTH1）. In the in vitro experiment， primary stromal cells were isolated from ectopic lesions of EMs patients. Cell counting kit-8 （CCK-8） was used to determine the optimal concentration of drug-containing serum for intervention. The level of reactive oxygen species （ROS） was measured， and Real-time PCR was applied to detect ferroptosis-related indicators. In the animal experiments， an EM rat model was established， and the rats were randomly assigned to the sham operation group， EMs group， low-dose Luoshi Neiyi Formula group （7.87 g·kg^-1）， high-dose Luoshi Neiyi prescription group （15.74 g·kg^-1）， and danazol group （42 mg·kg^-1）. Untargeted metabolomics detection and pathway enrichment analysis were conducted on serum samples from patients and rats. Spearman correlation analysis was performed to assess the relationship between differential metabolites and ferroptosis indicators. The correlations between differential metabolites in patient endometrium and serum and key ferroptosis indicators （GPX4， Fe²⁺， MDA， GSH） as well as ferroptosis-related mRNAs （GPX4， SLC7A11， FTH1） were analyzed， and correlation heatmaps were generated accordingly. ResultsCompared with normal eutopic endometrium， ectopic lesions in EMs patients showed glandular disorganization and stromal fibrosis. In ectopic endometrium， the contents of MDA， ROS， and Fe²⁺ decreased， while GPX4 level increased， and the mRNA expression of SLC7A11 and GPX4 was upregulated （P<0.05， P<0.01）. In serum， the levels of GPX4 and Fe²⁺ were elevated， whereas the GSH level declined， suggesting abnormalities in ferroptosis-related pathways in ectopic lesions （P<0.05， P<0.01）. After intervention with Luoshi Neiyi prescription-containing serum， the intracellular ROS level in ectopic endometrial stromal cells was elevated， the mRNA expression of SLC7A11 and GPX4 was downregulated， and TFR mRNA expression was upregulated （P<0.05， P<0.01）. Metabolomics analysis revealed 1104 and 198 differential metabolites in EMs patients and EMs rats， respectively， compared with their corresponding control groups， and both low-dose and high-dose Luoshi Neiyi prescription were found to regulate this metabolic disturbance， with the core regulatory pathways mainly involving arginine and proline metabolism. Correlation analysis showed that the glycerophospholipids including PI（16∶0/17∶0） and PI［18∶2（9Z，12Z）］ were negatively correlated with GPX4 and positively correlated with MDA， while 17α-hydroxyprogesterone was positively correlated with GPX4， SLC7A11， and FTH1 q<0.05）. ConclusionLuoshi Neiyi prescription may systematically ameliorate disease-associated metabolic dysregulation via modulation of the serum arginine and proline metabolism pathway， and may regulate ferroptosis in ectopic lesions through a mechanism potentially linked to the serum glycerophospholipid and steroid metabolism pathways. Collectively， these findings provide experimental evidence for the clinical application of Luoshi Neiyi prescription.

ObjectiveThis study aimed to investigate the mechanism by which Luoshi Neiyi prescription treats endometriosis （EMs） through regulating ferroptosis， and to screen key metabolites and analyze their association with ferroptosis. MethodsClinical samples of normal endometrium from patients without EMs and eutopic and ectopic endometrium from EMs patients （10 cases each） were collected and divided into control group， eutopic group， and EMs group. Hematoxylin-eosin （HE） staining was performed to observe ectopic lesions of EMs. Immunohistochemistry was used to detect the expression of solute carrier family 7 member 11 （SLC7A11） and glutathione peroxidase 4 （GPX4）. Enzyme-linked immunosorbent assay （ELISA） was adopted to determine the levels of malondialdehyde （MDA）， ferrous ion （Fe²⁺）， GPX4 and glutathione （GSH） in endometrial tissues， as well as serum levels of Fe²⁺， GPX4 and GSH. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of SLC7A11， GPX4， transferrin receptor （TFR） and ferritin heavy chain 1 （FTH1）. In the in vitro experiment， primary stromal cells were isolated from ectopic lesions of EMs patients. Cell counting kit-8 （CCK-8） was used to determine the optimal concentration of drug-containing serum for intervention. The level of reactive oxygen species （ROS） was measured， and Real-time PCR was applied to detect ferroptosis-related indicators. In the animal experiments， an EM rat model was established， and the rats were randomly assigned to the sham operation group， EMs group， low-dose Luoshi Neiyi Formula group （7.87 g·kg^-1）， high-dose Luoshi Neiyi prescription group （15.74 g·kg^-1）， and danazol group （42 mg·kg^-1）. Untargeted metabolomics detection and pathway enrichment analysis were conducted on serum samples from patients and rats. Spearman correlation analysis was performed to assess the relationship between differential metabolites and ferroptosis indicators. The correlations between differential metabolites in patient endometrium and serum and key ferroptosis indicators （GPX4， Fe²⁺， MDA， GSH） as well as ferroptosis-related mRNAs （GPX4， SLC7A11， FTH1） were analyzed， and correlation heatmaps were generated accordingly. ResultsCompared with normal eutopic endometrium， ectopic lesions in EMs patients showed glandular disorganization and stromal fibrosis. In ectopic endometrium， the contents of MDA， ROS， and Fe²⁺ decreased， while GPX4 level increased， and the mRNA expression of SLC7A11 and GPX4 was upregulated （P<0.05， P<0.01）. In serum， the levels of GPX4 and Fe²⁺ were elevated， whereas the GSH level declined， suggesting abnormalities in ferroptosis-related pathways in ectopic lesions （P<0.05， P<0.01）. After intervention with Luoshi Neiyi prescription-containing serum， the intracellular ROS level in ectopic endometrial stromal cells was elevated， the mRNA expression of SLC7A11 and GPX4 was downregulated， and TFR mRNA expression was upregulated （P<0.05， P<0.01）. Metabolomics analysis revealed 1104 and 198 differential metabolites in EMs patients and EMs rats， respectively， compared with their corresponding control groups， and both low-dose and high-dose Luoshi Neiyi prescription were found to regulate this metabolic disturbance， with the core regulatory pathways mainly involving arginine and proline metabolism. Correlation analysis showed that the glycerophospholipids including PI（16∶0/17∶0） and PI［18∶2（9Z，12Z）］ were negatively correlated with GPX4 and positively correlated with MDA， while 17α-hydroxyprogesterone was positively correlated with GPX4， SLC7A11， and FTH1 q<0.05）. ConclusionLuoshi Neiyi prescription may systematically ameliorate disease-associated metabolic dysregulation via modulation of the serum arginine and proline metabolism pathway， and may regulate ferroptosis in ectopic lesions through a mechanism potentially linked to the serum glycerophospholipid and steroid metabolism pathways. Collectively， these findings provide experimental evidence for the clinical application of Luoshi Neiyi prescription.

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

Objective To observe the analgesic and rehabilitation effects of non μ-opioids anesthesia/analgesia(NΜOA)based on quadratus lumborum block(QLB)in emergency cesarean section under general anesthesia.Methods The retrospective study method was adopted,50 pregnant women undergoing hysterectomy under emergency general anesthesia in Langfang People's Hospital from January 2023 to December 2024 were selected as the study objects.The patients were divided into μ-opioids anesthesia/analgesia(ΜOA)group and NΜOA group according to different anesthesia/analgesia methods,25 cases in each group.ΜOA group received ΜOA;NΜOA group received NΜOA+QLB.Incisional pain and uterine contraction pain numerical rating scale(NRS)at out of the post-anesthesia care unit(T1),intravenous injection of oxytocin(T2),press the palace bottom 24 hours(T3),out of bed activity after operation(T4)and first analgesic time of incision pain,first analgesic time of uterine contraction pain,first no vomiting eating time,first exhaust time was observed and recorded.The incidence of vasoactive agents during the anesthetic period,rescue analgesia,rescue antiemetic,constipation,sleep disturbance after operation within 48 hours after operation were also recorded.Results The NRS scores at T1,T2,T3 and T4 in ΜOA group were significantly higher than those in NΜOA group(incisional pain 3.36±1.25 vs.1.12±0.97,3.68±1.18 vs.2.00±0.91,5.76±1.67 vs.4.20±1.00,4.48±1.29 vs.3.32±0.95;uterine contraction pain 3.72±1.49 vs.1.24±1.05,4.64±1.60 vs.3.04±1.27,7.56±1.71 vs.5.16±1.37,3.56±0.22 vs.2.56±0.16,all P<0.05).The first analgesic time of incision pain,first analgesic time of uterine contraction pain in ΜOA group were significantly less than that in NΜOA group(hours:3.06±2.02 vs.17.48±10.93,2.68±2.22 vs.15.80±11.39,both P<0.05),the first no vomiting eating time,first exhaust time in ΜOA group were significantly longer than those in NΜOA group(hours:8.56±0.57 vs.6.32±0.14,15.44±1.42 vs.10.16±1.14,both P<0.05),the incidence of vasoactive agents,rescue analgesia,rescue antiemetic,constipation,sleep disturbance after operation within 48 hours in ΜOA group were significantly higher than those in NΜOA group[64.0%(16/25)vs.32.0%(8/25),48.0%(12/25)vs.20.0%(5/25),44.0%(11/25)vs.16.0%(4/25),64.0%(16/25)vs.36.0%(9/25),60.0%(15/25)vs.32.0%(8/25),all P<0.05].Conclusion NΜOA based on QLB safely and effectively reduced side effects of μ-opioids and enhanced recovery compared to ΜOA on emergency cesarean section patients undergoing general anesthesia.

Objective:To investigate the application effect of quadratus lumborum block (QLB) combined with less opioid anesthesia analgesia in laparoscopic total hysterectomy (LTH) in elderly patients with uterine prolapse.Methods:A total of 86 elderly patients with uterine prolapse who received LTH in the Langfang People’s Hospital from Jan. 2021 to Oct. 2023 were prospectively selected and divided into group A ( n=42) and group B ( n=44) according to the random number table method. Group A received transversus abdominis block (TAPB) + general anesthesia, and group B received QLB combined with less opioid anesthesia + general anesthesia. The pain duration at different time points after operation [Numeric Rating Scale (NRS) method to evaluate resting incision and visceral pain] was compared between the two groups. The dosage of opioids and the additional analgesia rate were compared between the two groups. The sleep quality of the two groups before and 72 hours after operation was compared, and the adverse reactions of anesthesia were counted. Results:Compared with 6 h after operation, NRS scores of resting incision pain and visceral pain in the two groups decreased gradually at 12, 24 and 48 h after operation, and it was lower in group B than in group A. There were statistically significant differences between groups ( P<0.05) . Dosage of sufentanil and remifentanil in group B was lower than that in group A, and the postoperative additional analgesia rate in group B (9.09%) was lower than that in group A (28.57%) ( P<0.05) . 72 hours after surgery, the sleep efficiency and total sleep time of patients in both groups were lower than those before surgery, and they were higher in group B than in group A; The awakening time and number of awakenings in both groups were higher than those before surgery, while they were lower in group B than in group A ( P<0.05) . The total incidence of adverse reactions in group B was 4.76% (2/44) , lower than 23.81% (10/42) of group A ( P<0.05) . Conclusion:QLB combined with less opioid anesthesia analgesia can effectively reduce the degree of incision and visceral pain after LTH in elderly patients with uterine prolapse, reduce the amount of opioids used during operation and the rate of additional analgesia, and reduce the incidence of adverse reactions of anesthesia.

Objective:To investigate the application effect of quadratus lumborum block (QLB) combined with less opioid anesthesia analgesia in laparoscopic total hysterectomy (LTH) in elderly patients with uterine prolapse.Methods:A total of 86 elderly patients with uterine prolapse who received LTH in the Langfang People’s Hospital from Jan. 2021 to Oct. 2023 were prospectively selected and divided into group A ( n=42) and group B ( n=44) according to the random number table method. Group A received transversus abdominis block (TAPB) + general anesthesia, and group B received QLB combined with less opioid anesthesia + general anesthesia. The pain duration at different time points after operation [Numeric Rating Scale (NRS) method to evaluate resting incision and visceral pain] was compared between the two groups. The dosage of opioids and the additional analgesia rate were compared between the two groups. The sleep quality of the two groups before and 72 hours after operation was compared, and the adverse reactions of anesthesia were counted. Results:Compared with 6 h after operation, NRS scores of resting incision pain and visceral pain in the two groups decreased gradually at 12, 24 and 48 h after operation, and it was lower in group B than in group A. There were statistically significant differences between groups ( P<0.05) . Dosage of sufentanil and remifentanil in group B was lower than that in group A, and the postoperative additional analgesia rate in group B (9.09%) was lower than that in group A (28.57%) ( P<0.05) . 72 hours after surgery, the sleep efficiency and total sleep time of patients in both groups were lower than those before surgery, and they were higher in group B than in group A; The awakening time and number of awakenings in both groups were higher than those before surgery, while they were lower in group B than in group A ( P<0.05) . The total incidence of adverse reactions in group B was 4.76% (2/44) , lower than 23.81% (10/42) of group A ( P<0.05) . Conclusion:QLB combined with less opioid anesthesia analgesia can effectively reduce the degree of incision and visceral pain after LTH in elderly patients with uterine prolapse, reduce the amount of opioids used during operation and the rate of additional analgesia, and reduce the incidence of adverse reactions of anesthesia.

Objective To observe the analgesic and rehabilitation effects of non μ-opioids anesthesia/analgesia(NΜOA)based on quadratus lumborum block(QLB)in emergency cesarean section under general anesthesia.Methods The retrospective study method was adopted,50 pregnant women undergoing hysterectomy under emergency general anesthesia in Langfang People's Hospital from January 2023 to December 2024 were selected as the study objects.The patients were divided into μ-opioids anesthesia/analgesia(ΜOA)group and NΜOA group according to different anesthesia/analgesia methods,25 cases in each group.ΜOA group received ΜOA;NΜOA group received NΜOA+QLB.Incisional pain and uterine contraction pain numerical rating scale(NRS)at out of the post-anesthesia care unit(T1),intravenous injection of oxytocin(T2),press the palace bottom 24 hours(T3),out of bed activity after operation(T4)and first analgesic time of incision pain,first analgesic time of uterine contraction pain,first no vomiting eating time,first exhaust time was observed and recorded.The incidence of vasoactive agents during the anesthetic period,rescue analgesia,rescue antiemetic,constipation,sleep disturbance after operation within 48 hours after operation were also recorded.Results The NRS scores at T1,T2,T3 and T4 in ΜOA group were significantly higher than those in NΜOA group(incisional pain 3.36±1.25 vs.1.12±0.97,3.68±1.18 vs.2.00±0.91,5.76±1.67 vs.4.20±1.00,4.48±1.29 vs.3.32±0.95;uterine contraction pain 3.72±1.49 vs.1.24±1.05,4.64±1.60 vs.3.04±1.27,7.56±1.71 vs.5.16±1.37,3.56±0.22 vs.2.56±0.16,all P<0.05).The first analgesic time of incision pain,first analgesic time of uterine contraction pain in ΜOA group were significantly less than that in NΜOA group(hours:3.06±2.02 vs.17.48±10.93,2.68±2.22 vs.15.80±11.39,both P<0.05),the first no vomiting eating time,first exhaust time in ΜOA group were significantly longer than those in NΜOA group(hours:8.56±0.57 vs.6.32±0.14,15.44±1.42 vs.10.16±1.14,both P<0.05),the incidence of vasoactive agents,rescue analgesia,rescue antiemetic,constipation,sleep disturbance after operation within 48 hours in ΜOA group were significantly higher than those in NΜOA group[64.0%(16/25)vs.32.0%(8/25),48.0%(12/25)vs.20.0%(5/25),44.0%(11/25)vs.16.0%(4/25),64.0%(16/25)vs.36.0%(9/25),60.0%(15/25)vs.32.0%(8/25),all P<0.05].Conclusion NΜOA based on QLB safely and effectively reduced side effects of μ-opioids and enhanced recovery compared to ΜOA on emergency cesarean section patients undergoing general anesthesia.

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

Loss-of-function variants in CSDE1 have been strongly linked to neuropsychiatric disorders, yet the precise role of CSDE1 in neurogenesis remains elusive. In this study, we demonstrate that knockout of Csde1 during cortical development in mice results in impaired neural progenitor proliferation, leading to abnormal cortical lamination and embryonic lethality. Transcriptomic analysis revealed that Csde1 upregulates the transcription of genes involved in the cell cycle network. Applying a dual thymidine-labelling approach, we further revealed prolonged cell cycle durations of neuronal progenitors in Csde1-knockout mice, with a notable extension of the G1 phase. Intersection with CLIP-seq data demonstrated that Csde1 binds to the 3' untranslated region (UTR) of mRNA transcripts encoding cell cycle genes. Particularly, we uncovered that Csde1 directly binds to the 3' UTR of mRNA transcripts encoding Cdk6, a pivotal gene in regulating the transition from the G1 to S phases of the cell cycle, thereby maintaining its stability. Collectively, this study elucidates Csde1 as a novel regulator of Cdk6, sheds new light on its critical roles in orchestrating brain development, and underscores how mutations in Csde1 may contribute to the pathogenesis of neuropsychiatric disorders.
Animals ; Neurogenesis/genetics* ; Cell Cycle/genetics* ; Mice, Knockout ; Mice ; Neural Stem Cells/metabolism* ; DNA-Binding Proteins/metabolism* ; Cyclin-Dependent Kinase 6/genetics* ; Cell Proliferation ; 3' Untranslated Regions ; Cerebral Cortex/embryology* ; RNA-Binding Proteins ; Mice, Inbred C57BL

Background Air pollution remains a critical public health issue, with persistent exposure to air pollutants continuing to pose significant health risks. Currently, research investigating the association between air pollution and myocardial infarction mortality in Shenzhen remains inadequate. Objective To quantitatively assess the association between air pollutants and myocardial infarction mortality in residents. Methods Based on the mortality surveillance system of Shenzhen Center for Disease Control and Prevention, we conducted a time-stratified case-crossover study of 10089 permanent residents who died from myocardial infarction in Shenzhen between 2013 and 2022. Using residential address information, we obtained individual-level exposure data for air pollutants from the China High Air Pollutants dataset and meteorological factors from the China Meteorological Administration Land Data Assimilation System. A time-stratified case-crossover study design was employed to construct a conditional logistic regression model to assess the association between short-term exposure to air pollutants and myocardial infarction mortality. The exposure-response relationship was visualized, and a comprehensive health risk assessment was performed. Results This study included a total of 10 089 cases of myocardial infarction deaths in Shenzhen. The median [interquartile range (IQR)] concentrations of fine particulate matter (PM_2.5), inhalable particulate matter (PM₁₀), sulfur dioxide (SO₂), nitrogen dioxide (NO₂), carbon monoxide (CO), and ozone (O₃) in Shenzhen from 2013 to 2022 were 24.59 (20.19) μg·m⁻³, 42.85 (28.42) μg·m⁻³, 8.53 (3.39) μg·m⁻³, 29.47 (13.56) μg·m⁻³, 0.77 (0.27) mg·m⁻³, and 86.53 (51.39) μg·m⁻³, respectively. The moving average concentrations of PM_2.5 and PM₁₀ over the current day and the previous 2 days (lag02) showed the highest risk of myocardial infarction mortality, with an odds ratio (OR) and 95% confidence interval (95%CI) of 1.004 (1.001, 1.007) and 1.004 (1.002, 1.006), respectively. At lag05, SO₂ demonstrated the strongest association with the risk of myocardial infarction mortality, with an OR (95%CI) of 1.042 (1.019, 1.065). The exposure-response relationships of PM_2.5, PM₁₀, and SO₂ with myocardial infarction mortality were approximately linear, whereas NO₂ exhibited a nonlinear relationship. At lag05, the highest risk of myocardial infarction mortality associated with NO₂ exposure was observed when the NO₂ concentration was ≤21.92 μg·m⁻³, with an OR (95% CI) of 1.024 (1.003, 1.046). The health risk assessment indicated that, using the WHO Air Quality Guidelines as the reference, the local PM_2.5 and NO₂ exposure led to an excess mortality of 398 and 298 cases, respectively. The sensitivity analysis revealed that after adjusting for O₃ and restricting the study period to 2013—2019, the effect estimates for PM_2.5, PM₁₀, NO₂, and SO₂ on myocardial infarction mortality slightly increased. Conclusion Short-term exposure to air pollutants, including PM_2.5, PM₁₀, NO₂, and SO₂, could increase the risk of myocardial infarction mortality. This study provides important scientific evidence for environmental health risk assessment and environmental management in Shenzhen.