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.

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.

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.

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 explore the influencing factors for sarcopenia among elderly male patients with type 2 diabetes (T2DM), so as to provide the basis for the early prevention and treatment of sarcopenia. Methods: Male T2DM patients aged 60 and above admitted to Hangzhou First People's Hospital from January to December 2024 were selected as the study subjects. Demographic data, T2DM complications, and blood biochemical parameters were collected. Physical activity levels were assessed using the Chinese version of the International Physical Activity Questionnaire. The diagnosis of sarcopenia was made according to the diagnostic procedures and criteria established by the Asian Working Group for Sarcopenia in 2019. Factors affecting sarcopenia among elderly male patients with T2DM were analyzed using multivariable logistic regression model. Results: A total of 455 elderly male patients with T2DM were surveyed, with a mean age of (71.80±9.55) years. The predominant physical activity level was moderate with 226 cases accounting for 49.67%. The disease course of T2DM was mainly from 10-<20 years, with 229 cases accounting for 50.33%. There were 140 cases of T2DM complications, accounting for 30.77%. A total of 138 cases of sarcopenia were detected, with a prevalence of 30.33%. Multivariable logistic regression analysis showed that age (OR=1.077, 95%CI: 1.003~1.156), body mass index (<18.5kg/m2, OR=11.056, 95%CI: 3.343~36.547; 18.5~<25.0 kg/m2, OR=2.633, 95%CI: 1.420~4.881), physical activity level (low, OR=2.469, 95%CI: 1.421~4.292), chronic obstructive pulmonary disease (yes, OR=1.871, 95%CI: 1.091~3.206), T2DM complications (yes, OR=3.015, 95%CI: 1.516~6.001), glycated hemoglobin (≥7%, OR=2.822, 95%CI: 1.423~5.590) and albumin (OR=0.810, 95%CI: 0.662~0.991) were factors affecting sarcopenia among elderly male patients with T2DM (P<0.05). Conclusion Advanced age, body mass index <25.0 kg/m2, low physical activity level, chronic obstructive pulmonary disease, T2DM complications, high glycated hemoglobin and low albumin are associated with a higher risk of sarcopenia in elderly male patients with T2DM.

Objective To observe the effects of folic acid(FA)supplementation on the expression of Flotillin-1 and β-amyloid protein(Aβ)-metabolism-related proteins in the brains of inflammation-stimulated Alzheimer's disease(AD)mice.Methods Twenty-seven 6-month-old male APP/PS1 mice were randomly divided into AD,AD+LPS,and AD+LPS+FA groups,with nine mice in each group.Nine C57BL/6J male mice born within the same month were used as the Control group.The AD+LPS+FA group was given folic-acid-supplemented feed(8 mg/kg)for 3 months of intervention,while the other three groups were fed normal feed.Lipopolysaccharide solution(LPS,250 μg/(kg·d))was injected intraperitoneally into mice in the AD+LPS and AD+LPS+FA groups 1 week before the end of the experiment,and saline was injected into the remaining two groups.The serum inflammatory factors TNF-α and IL-6 levels and brain tissue Aβ1-40 and Aβ1-42 levels of mice in each group were detected by ELISA.Flotillin-1 protein expression in brain tissue was detected using Western blot,and the co-expression of Flotillin-1 and Aβ1-42/APP/PS1/BACE1 in the cortical region of the brain was detected via immunofluorescence double-labeling.Results After ANOVA analysis,we found mice in the AD group had elevated serum TNF-α and IL-6 levels(P＜0.05),elevated levels of Aβ1-40 and Aβ1-42(P＜0.05),increased expression of Flotillin-1 protein(P＜0.05),and increased co-expression of Flotillin-1 and Aβ1-42/APP/PS1/BACE1 in the cortical brain tissue(P＜0.05)compared with the Control group.Compared with mice in the AD group,those in the AD+LPS group had further increases in serum inflammatory factors and Aβ levels in the brain(P＜0.05)and increased co-expression of Flotillin-1 and Aβ1-42/APP/BACE1 double-labeled proteins in their cortical brain tissue(P＜0.05).Compared with mice in the AD+LPS group,those in the AD+LPS+FA group had lower in vivo inflammation levels and Aβ content in the brain(P＜0.05),lower brain tissue Flotillin-1 protein expression(P＜0.05),and lower Flotillin-1 and Aβ1-42/APP/PS1/BACE1 protein co-expression in cortical brain tissue(P＜0.05).Conclusions Folic acid supplementation may reduce Flotillin-1 protein expression and Aβ deposition in the brain of AD inflammatory mice.

Objective To explore the pathological features of lung tissue in mild chronic obstructive pulmonary disease(COPD)and their association with inflammatory factors.Methods A total of 70 patients who underwent surgery for small lung nodule were prospectively included,and were divided into the normal group(n=10),the mild COPD group(n=50)and the moderate and severe COPD group(n=10).The pathological changes of lung tissue were evaluated after HE,Masson and EVG staining.The expression levels of SMA,Actin and CD31 proteins were detected by immunohistochemistry staining.Tumor necrosis factor α(TNF-α),interleukin-10(IL-10)protein and mRNA levels were detected by immunohistochemistry and qPCR.Results Pulmonary tissue in mild COPD showed widening of alveolar septum,dilation of small airways,mild thickening of blood vessel wall and inflammatory reaction dominated by lymphocyte infiltration.Immunohistochemistry staining showed that contents of SMA and Actin proteins in mild COPD lung tissue were higher than those in the normal group(P＜0.05).In addition,the TNF-α mRNA and the positive rate of TNF-α in lung tissue of mild COPD were significantly higher than those in the normal group,while the IL-10 mRNA was significantly lower than that of the normal group(all P＜0.05).SMA and Actin were positively correlated with the positive expression of inflammatory cytokine TNF-α,but negatively correlated with the positive expression of IL-10(all P＜0.05).Conclusion The main pathological changes of lung tissue in mild COPD include small lung blood vessel remodeling ocharacterized by thickening of small blood vessel smooth muscle layer and lymphocyte-dominated inflammatory response,while the increase of pro-inflammatory factor TNF-α and decrease of anti-inflammatory factor IL-10 are associated with pathological changes of COPD.

AIM:To investigate the effects of the compound ANBP on wound healing in diabetic rats and ex-plore its mechanism of action.METHODS:Ninety male SD rats were randomly divided into blank,model,compound ANBP,Beifuxin,and nicotinamide mononucleotide(NMN)groups,with 16 rats in each group.Wound healing in each group was observed and samples were taken on days 3,7 and 14 to analyze the wound healing rate.Local histopathological changes were observed using HE and Masson staining.The expressions of pyruvate dehydrogenase E1 subunit alpha 1(PDHA1),citrate synthase(CS),isocitrate dehydrogenase(IDH1)and oxoglutarate dehydrogenase(OGDH)were de-tected through immunofluorescence and Western blot.The number and morphology of mitochondria in the wound tissue were observed using transmission electron microscopy.RESULTS:Histomorphological changes revealed significant im-provement in diabetic wound healing in the blank and compound ANBP groups compared to that of the model group.The wound healing rates of the blank,compound ANBP,Beifuxin,and NMN groups were significantly increased on days 3,7,and 14(P<0.01).Compared to the model group,granulation tissue generation was higher in the other groups,cover-ing the wound defect and producing abundant collagen fibers.At 3,7,and 14 days after intervention,the blank,com-pound ANBP,Beifuxin,and NMN groups showed significantly enhanced fluorescence intensities of TCA cycling-related enzymes PDHA1,CS,IDH1,and OGDH indicating increased expression of these enzymes.The levels of the TCA cy-cling-related enzymes were significantly increased(P<0.01)in the compound ANBP,Beifuxin and NMN groups but were significantly decreased(P<0.01)in the model group.An increase in the number and density of mitochondria and a de-crease in the cavitation rate of mitochondria with improved morphology(P<0.05)was observed in the group treated with compound ANBP.CONCLUSION:Compound ANBP may increase the number of mitochondria,improve mitochondrial morphology and function,upregulate the expression levels of PDHA1,CS,IDH1,and OGDH proteins,and accelerate the regeneration of wound granulation tissue,thus promoting the healing of diabetic wounds in rats.

Objective:To study the effect of tertiary lymphoid structures(TLS)on the pathological response and prognosis of patients with non-small cell lung cancer(NSCLC)receiving neoadjuvant therapy.Methods:We retrospectively collected the data of 132 patients with NSCLC who underwent neoadjuvant therapy and surgery at Tianjin Chest Hospital between January 2019 and December 2023,including 40 in the neoadjuvant chemotherapy(NC)group and 92 in the NC plus immunotherapy(NCI)group.The percentage of residual viable tumor(RVT)and tumor infiltrating lymphocyte(TIL)counts were evaluated by hematoxylin and eosin(H&E)staining,while TLS number and matur-ity were assessed by H&E and immunohistochemical staining.The differences in TLS number and maturity and effects on patient pathologic-al response and prognosis were compared between groups.Results:TIL count,total TLS number,pathological complete response and major pathological response rates were significantly higher in the NCI versus NC group(P<0.001).Moreover,a multivariate Logistic analysis sho-wed that TLS number and maturity and TIL count affected pathological response in the NCI group(P<0.05).A multiple linear regression ana-lysis indicated that a low TIL count was a risk factor for a high RVT in the NC group,while a low number of mature TLS,low TIL count,and N stage were independent risk factors for a high RVT in the NCI group(all P<0.05).In the NCI group,a multivariate Cox regression analysis showed that a low number of mature TLS(P=0.001)and low TIL count(P=0.009)were independent predictors of disease-free survival(DFS),while a survival analysis showed that patients in the NCI group with high(vs.low)numbers of mature TLS and a high(vs.low)TIL count had significantly longer DFS(all P<0.001).Conclusions:A low number of mature TLS and low TIL count were associated with an adverse patholo-gical response and short DFS in patients with NSCLC.Thus,TLS maturity and TIL count can predict the pathological response and prognosis of patients with NSCLC treated with NCI.

Objective To study the effects of ultrasound-guided quadratus lumborum block(QLB)on intraoperative hemodynamics and opioid dosage in emergency patients with ectopic pregnancy.Methods A total of 70 patients with ectopic pregnancy undergoing laparoscopic surgery in Langfang People's Hospital from January 2021 to February 2024 were selected as subjects.According to the different anesthesia methods,the patients were divided into the control group and the study group,with 35 cases in each group.The control group was given general anesthesia,while the study group additionally added ultrasound-guided QLB.The intraoperative sedation effect,hemodynamics,postoperative pain,incidence of adverse reactions and opioid use at different times(admission,entry,intubation,skin incision,extubation,and discharge)were observed in the two groups.Results There were no statistically significant differences in the onset time of sedation,the rate of salvage sedation,the incidence of intraoperative body movements,the modified observer's assessment of alert/sedation(MOAA/S)at each time,and the hemodynamics at the time of admission,entry and intubation between the two groups.The mean arterial pressure(MAP),systolic blood pressure(SBP)and heart rate(HR)in the study group were significantly lower than those in the control group during skin incision,extubation and discharge[skin incision:MAP(mmHg,1 mmHg≈0.133 kPa)was 85.24±4.59 vs.96.95±4.68,SBP(mmHg)was 92.24±4.85 vs.99.49±5.13,HR(times/min)was 85.33±2.96 vs.94.51±2.92;extubation:MAP(mmHg)was 94.84±5.02 vs.102.05±5.13,SBP(mmHg)was 96.48±4.72 vs.105.03±5.07,HR(times/min)was 95.51±4.95 vs.102.49±5.87;discharge:MAP(mmHg)was 86.14±4.99 vs.93.71±5.25,SBP(mmHg)was 96.48±4.69 vs.104.37±5.02,HR(times/min)was 84.05±4.57 vs.90.51±4.86,all P<0.05]and pulse oxygen saturation(SpO2)was higher than those in the control group(skin incision:0.988 5±0.012 2 vs.0.965 4±0.012 3,extubation:0.974 7±0.012 4 vs.0.963 2±0.012 1,discharge:0.981 1±0.012 4 vs.0.970 3±0.012 3,all P<0.05).The resting numeric rating scale(NRS)scores and active NRS scores in the study group were lower than those in the control group at 3,6,12,and 24 hours after surgery,the random time was prolonged,the resting NRS and active NRS in the two groups gradually increased,reaching a peak at 24 hours after surgery,and the resting NRS and active NRS in the study group were significantly lower than those in the control group(resting NRS:3.86±0.82 vs.4.53±1.04,active NRS:4.26±1.05 vs.4.85±1.13,all P<0.05).The incidence of adverse reactions in the study group was lower than that in the control group[11.43%(4/35)vs.34.29%(12/35),P<0.05].The dosage of Sufentanil in 24 hours and 48 hours,the number of analgesic pump in 48 hours and the number of relief analgesia cases in the study group were lower than those in the control group[the dosage of Sufentanil in 24 hours(μg):23.28±4.02 vs.36.14±4.57,the dosage of Sufentanil in 48 hours(μg):41.61±4.82 vs.59.33±6.25,the number of analgesic pump in 48 hours(times):2.94±1.22 vs.6.15±1.71,the proportion of relief analgesia:8.57%(3/35)vs.28.57%(10/35),all P<0.05].Conclusion Ultrasound-guided QLB can reduce hemodynamic fluctuations,relieve postoperative pain,reduce adverse reactions and opioid use in emergency patients with ectopic pregnancy,demonstrating a positive impact.