Background: The treatment landscape for melanoma, a particularly malignant skin cancer, is constrained by notable drug resistance and toxicity. β-Asarone, a natural compound from Acorus tatarinowii, has shown anticancer potential. Disruption of calcium homeostasis and mitochondrial dysfunction are key regulators of tumor cell survival and death. Objective: This research was conducted to investigate the impact of β-Asarone on B16F10 melanoma cells, focusing on its potential to induce apoptosis by modulating calcium signaling and mitochondrial function. Methods: Cell proliferation and apoptosis were evaluated using CCK-8, colony formation, EdU, and TUNEL assays. Intracellular calcium levels and mitochondrial membrane potential were measured using Fluo-4 AM, Rhod-2 AM, and JC-1 staining. Reactive oxygen species (ROS) generation and adenosine triphosphate (ATP) levels were assessed by fluorescent probes and ATP assay. Western blotting was utilized to detect apoptosis-related proteins, AMP-activated protein kinase (AMPK) pathway activation, and mitochondrial dynamics (OPA1, DRP1, FIS1). Results: Treatment with β-Asarone notably inhibited the proliferation of B16F10 cells while simultaneously inducing apoptosis. Fluorescent probe analysis revealed that β-Asarone triggered cytosolic and mitochondrial Ca 2+ overloaded in both the cytosol and mitochondria, accompanied by decreased mitochondrial membrane potential, elevated ROS levels, and reduced ATP production. Western blot analysis showed increased expression of DRP1 and FIS1, decreased OPA1, and enhanced AMPK phosphorylation, indicating that β-Asarone promotes mitochondrial fission through AMPK activation, likely driven by intracellular calcium imbalance. Conclusion This study demonstrates that β-Asarone induces apoptosis in B16F10 melanoma cells by triggering Ca 2+ overload and mitochondrial dysfunction.

Objective:To investigate the effects of melatonin(Mel)on inflammatory damage in the retina of rats with oxygen-induced retinopathy(OIR)and the molecular mechanisms.Methods:Healthy neonatal SD rats were di-vided into the sham group(Sham),the model group(OIR),and the melatonin treatment group(OIR+Mel).The OIR model was induced by alternating 50%/10%oxygen concentration exposure for 14 d.The OIR+Mel group was in-jected intraperitoneally with 10 mg-kg-1 melatonin.Hematoxylin-eosin(HE)staining was used to observe the morpho-logical changes in the retinal tissue;immunohistochemical staining was used to detect the expression of retinal cleaved-caspase-1 and IL-1β proteins;and immunofluorescence staining was used to detect the expression of cGAS-STING-NL-RP3 signaling molecules and gasdermin(GSDMD)in the microglia of the retina.Results:HE staining results showed that compared with the Sham group,the retinal cells in the OIR group were disorganized and the thickness of the inner retina was significantly thinner,and the retinal cells in the OIR+Mel group were more neatly arranged compared with those in the OIR group(P<0.05).Immunohistochemical staining results showed that the number of cleaved-caspase-1+and IL-1β+cells in the retina of rats in the OIR group increased significantly compared with that in the Sham group,and the number of cleaved-caspase-1+and IL-1β+cells in the retina of rats in the OIR+Mel group decreased signifi-cantly compared with that of the OIR group(P<0.05).Immunofluorescence staining results showed that the number of cGAS+,STING+and NLRP3+cells in the retina of rats in the OIR group increased significantly compared with that in the Sham group,and the number of cGAS+,STING+and NLRP3+cells in the retina of rats in the OIR+Mel group de-creased significantly compared with that in the OIR group(P<0.05);The number of Iba-1+/N-GSDMD+cells in-creased significantly in the OIR group compared with the Sham group,whereas the number of Iba-1+/N-GSDMD+cells in the OIR+Mel group was significantly less than that in the OIR group,but still more than that in the Sham group(P<0.05).Conclusion:Mel inhibits the pyroptosis of retinal microglia,thus attenuates retinal inflammatory injury in OIR rats,and its mechanism may be related to the cGAS-STING-NLRP3 signaling pathway.

Objective To explore the mechanism of the thioredoxin-interacting protein(TXNIP)/thioredoxin-1(Trx-1)pathway in regulating the polarization of retinal microglia in rats after retinal ischemia-reperfusion(RIR)in rats,and to provide new ideas for the prevention and treatment of retinal ischemia reperfusion injury(RIRI).Methods For-ty-two healthy adult male Sprague-Dawley rats were randomly divided into a Sham group,a RIRI group and a TXNIP siRNA group.The right eye of the rats was experimented.For RIRI and TXNIP siRNA groups,RIRI models were established using the anterior chamber high intraocular pressure method.Rats in the TXNIP siRNA group were given the intravitreal injection of TXNIP siRNA 3 d before modeling.Hematoxylin-eosin(HE)staining was used to analyze retinal histopathologic changes of rats in all groups 24 h after modeling.Immunohistochemical staining of brain-specific homeobox/POU domain proteins 3A(Brn-3a)was made to count the number of retinal ganglion cells(RGCs).The dynamical changes in the number of TXNIP+cells 6 h,24 h,72 h and 7 d after modelling were analyzed through immunohistochemical staining in the RIRI group.The retinal microglia polarization and changes in the expression of TXNIP and Trx-1 proteins in each group were de-tected by double immunofluorescence staining and Western blot 24 h after modeling.Results HE staining results showed that 24 h after modelling,the retinal cells were disordered and the inner retinal layer was thickened and swelled in RIRI and TXNIP siRNA groups,compared with those in the Sham group(all P＜0.05).Immunohistochemical staining results of Brn-3a showed that 24 h after modeling,the number of Brn-3a+cells in RIRI and TXNIP siRNA groups significantly decreased,compared with that in the Sham group(both P＜0.05).The number of Brn-3a+cells in the TXNIP siRNA group was signifi-cantly higher than that in the RIRI group(P＜0.05).Immunohistochemical staining results of TXNIP at different time points after modeling showed that the expression of TXNIP+proteins started to increase 6 h after modeling.The TXNIP+protein level reached a peak at 24 h and then decreased gradually.Western blot results revealed that 24 h after modeling,RIRI and TXNIP siRNA groups had significantly higher TXNIP levels and significantly lower Trx-1 levels than the Sham group(all P＜0.05).Compared with those in the RIRI group,the expression of TXNIP proteins was significantly lower and the expression of Trx-1 proteins was significantly higher in the TXNIP siRNA group(both P＜0.05).Double immunofluores-cence staining showed that 24 h after modeling,Iba1+/CD206+cells were significantly more and Iba1+/CD16+cells were significantly less in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).RIRI and TXNIP siRNA groups had significantly more Ibal+/TXNIP+cells and significantly less Iba1+/Trx-1+cells than the Sham group(both P＜0.05).The number of Iba1+/TXNIP+cells was significantly lower and the number of Iba1+/Trx-1+cells was significantly higher in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).Conclusion RIR activates the TXNIP/Trx-1 path-way to induce the activation of retinal microglia and regulate the polarization of microglia,thereby resulting in RIRI in rats.

Objective:To investigate the effects of melatonin(Mel)on inflammatory damage in the retina of rats with oxygen-induced retinopathy(OIR)and the molecular mechanisms.Methods:Healthy neonatal SD rats were di-vided into the sham group(Sham),the model group(OIR),and the melatonin treatment group(OIR+Mel).The OIR model was induced by alternating 50%/10%oxygen concentration exposure for 14 d.The OIR+Mel group was in-jected intraperitoneally with 10 mg-kg-1 melatonin.Hematoxylin-eosin(HE)staining was used to observe the morpho-logical changes in the retinal tissue;immunohistochemical staining was used to detect the expression of retinal cleaved-caspase-1 and IL-1β proteins;and immunofluorescence staining was used to detect the expression of cGAS-STING-NL-RP3 signaling molecules and gasdermin(GSDMD)in the microglia of the retina.Results:HE staining results showed that compared with the Sham group,the retinal cells in the OIR group were disorganized and the thickness of the inner retina was significantly thinner,and the retinal cells in the OIR+Mel group were more neatly arranged compared with those in the OIR group(P<0.05).Immunohistochemical staining results showed that the number of cleaved-caspase-1+and IL-1β+cells in the retina of rats in the OIR group increased significantly compared with that in the Sham group,and the number of cleaved-caspase-1+and IL-1β+cells in the retina of rats in the OIR+Mel group decreased signifi-cantly compared with that of the OIR group(P<0.05).Immunofluorescence staining results showed that the number of cGAS+,STING+and NLRP3+cells in the retina of rats in the OIR group increased significantly compared with that in the Sham group,and the number of cGAS+,STING+and NLRP3+cells in the retina of rats in the OIR+Mel group de-creased significantly compared with that in the OIR group(P<0.05);The number of Iba-1+/N-GSDMD+cells in-creased significantly in the OIR group compared with the Sham group,whereas the number of Iba-1+/N-GSDMD+cells in the OIR+Mel group was significantly less than that in the OIR group,but still more than that in the Sham group(P<0.05).Conclusion:Mel inhibits the pyroptosis of retinal microglia,thus attenuates retinal inflammatory injury in OIR rats,and its mechanism may be related to the cGAS-STING-NLRP3 signaling pathway.

Objective To explore the mechanism of the thioredoxin-interacting protein(TXNIP)/thioredoxin-1(Trx-1)pathway in regulating the polarization of retinal microglia in rats after retinal ischemia-reperfusion(RIR)in rats,and to provide new ideas for the prevention and treatment of retinal ischemia reperfusion injury(RIRI).Methods For-ty-two healthy adult male Sprague-Dawley rats were randomly divided into a Sham group,a RIRI group and a TXNIP siRNA group.The right eye of the rats was experimented.For RIRI and TXNIP siRNA groups,RIRI models were established using the anterior chamber high intraocular pressure method.Rats in the TXNIP siRNA group were given the intravitreal injection of TXNIP siRNA 3 d before modeling.Hematoxylin-eosin(HE)staining was used to analyze retinal histopathologic changes of rats in all groups 24 h after modeling.Immunohistochemical staining of brain-specific homeobox/POU domain proteins 3A(Brn-3a)was made to count the number of retinal ganglion cells(RGCs).The dynamical changes in the number of TXNIP+cells 6 h,24 h,72 h and 7 d after modelling were analyzed through immunohistochemical staining in the RIRI group.The retinal microglia polarization and changes in the expression of TXNIP and Trx-1 proteins in each group were de-tected by double immunofluorescence staining and Western blot 24 h after modeling.Results HE staining results showed that 24 h after modelling,the retinal cells were disordered and the inner retinal layer was thickened and swelled in RIRI and TXNIP siRNA groups,compared with those in the Sham group(all P＜0.05).Immunohistochemical staining results of Brn-3a showed that 24 h after modeling,the number of Brn-3a+cells in RIRI and TXNIP siRNA groups significantly decreased,compared with that in the Sham group(both P＜0.05).The number of Brn-3a+cells in the TXNIP siRNA group was signifi-cantly higher than that in the RIRI group(P＜0.05).Immunohistochemical staining results of TXNIP at different time points after modeling showed that the expression of TXNIP+proteins started to increase 6 h after modeling.The TXNIP+protein level reached a peak at 24 h and then decreased gradually.Western blot results revealed that 24 h after modeling,RIRI and TXNIP siRNA groups had significantly higher TXNIP levels and significantly lower Trx-1 levels than the Sham group(all P＜0.05).Compared with those in the RIRI group,the expression of TXNIP proteins was significantly lower and the expression of Trx-1 proteins was significantly higher in the TXNIP siRNA group(both P＜0.05).Double immunofluores-cence staining showed that 24 h after modeling,Iba1+/CD206+cells were significantly more and Iba1+/CD16+cells were significantly less in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).RIRI and TXNIP siRNA groups had significantly more Ibal+/TXNIP+cells and significantly less Iba1+/Trx-1+cells than the Sham group(both P＜0.05).The number of Iba1+/TXNIP+cells was significantly lower and the number of Iba1+/Trx-1+cells was significantly higher in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).Conclusion RIR activates the TXNIP/Trx-1 path-way to induce the activation of retinal microglia and regulate the polarization of microglia,thereby resulting in RIRI in rats.

Objective To investigate the current status of pelvic floor muscle training(PFMT)adherence for preventing low anterior resection syndrome(LARS)in rectal cancer patients with prophylactic ostomy and analyze its influencing factors.Methods A total of 247 patients who underwent preventive stoma reversal surgery after sphincter-preserving rectal cancer resection between January 2024 and February 2025 at 22 tertiary hospitals in the Beijing-Tianjin-Hebei region were recruited in this study.Their clinical data were collected through a general information questionnaire,a LARS Knowledge-Attitude-Practice and Needs questionnaire,a PFMT Self-Efficacy Scale,and a PFMT Adherence Questionnaire.Univariate analysis,correlation analysis,LASSO regression,and random forest importance ranking were applied to screen the variables,and multiple linear regression analysis was conducted to analyze the significant variables.Results The overall PFMT adherence score was 14.52±4.18 among the 247 patients.The random forest algorithm identified 7 key predictors when the minimum error was achieved at a λ value of 2.293.The top 7 variables in importance ranking were PFMT self-efficacy,total knowledge-attitude-practice(KAP)score,education level,primary caregiver,tumor location,stoma reversal time,and chemoradiotherapy.Multiple linear regression analysis revealed that PFMT self-efficacy(P<0.001,β=0.007,95%CI:0.004～0.009),total KAP score(P<0.001,β=0.052,95%CI:0.035～0.070),stoma reversal time(P<0.030,β=-0.539,95%CI:-1.025～-0.053),and chemoradiotherapy(P<0.045,β=-0.451,95%CI:0.010～0.892)were significant related factors of PFMT adherence(P<0.05).Conclusion PFMT adherence for LARS prevention is at a moderate level in rectal cancer patients with prophylactic ostomy.Key factors such as PFMT self-efficacy,total KAP score,stoma reversal time,and chemoradiotherapy are significantly correlated with PFMT adherence.

Objective To evaluate the clinical efficacy of platelet-rich plasma(PRP)in treat-ment of diabetic foot ulcers(DFU)using a meta-analysis approach.Methods Relevant literature on PRP for treatment of DFU was collected by searching databases including PubMed,Cochrane Library,Embase,Web of Science,NEJM,CNKI,Wanfang Data,VIP Database,and China Biology Medicine disc.Relevant literature on PRP for treatment of DFU was collected.The literature was read,and effect sizes were extracted.The extracted data were then subjected to a meta-analysis using RevMan 5.3 software.Results A total of 6 studies were included in this meta-analysis,involving 440 pa-tients.Among them,there were 208 patients in experimental group(PRP combined with conventional treatment)and 232 patients in control group(conventional treatment,blank control,or placebo treat-ment).The ulcer healing efficacy rate in the experimental group was higher than that in the control group(OR=1.29,95％CI,1.19 to 1.40,P＜0.05).The incidence of adverse reactions in the experimental group was lower than that in the control group(OR=0.33,95％CI,0.12 to 0.93,P=0.94).The healing time(MD=-14.37,95％CI,-23.12 to-5.62,P=0.001),thickness of granulation tissue on the ulcer surface(MD=1.60,95％CI,1.31-1.88,P＜0.000 01),coverage rate of granulation tissue on ulcer surface(MD=6.03,95％CI,3.79 to 8.26,P＜0.05),and level of vascular endothelial growth factor on the ulcer surface(MD=7.62,95％CI,1.57 to 13.67,P=0.01)in the experimental group were all superior to those in the control group.Con-clusion PRP treatment for DFU has the advantages of improving the ulcer healing efficacy rate,shortening the ulcer healing time,promoting the growth of granulation tissue on the ulcer surface,and reducing the incidence of adverse reactions.

This paper summarizes clinical insights on treating the syndrome of coexisting dryness and dampness in Sjögren's syndrome （SS） based on the "three methods for liver diseases". It is proposed that the core pathogenesis of this subtype lies in the liver's failure to regulate and disperse， leading to fluid retention and transformation into dryness； disharmony between wood （liver） and earth （spleen）， resulting in internal generation of dryness and dampness； and liver impairment affecting the kidney， eventually causing extreme dryness to transform into dampness. This syndrome is thus characterized by disharmony between the liver and spleen， as well as dysfunction between ti （character， 体） and yong （function， 用）. Drawing from the Guide to Clinical Case （《临证指南医案》）， the "three methods for liver diseases" are applied as follows，i.e. using pungent and dispersing herbs to regulate the liver and resolve qi stagnation； using sweet and gentle herbs to soothe the liver and address the root of concurrent dryness and dampness； and using sour and purgative herbs to soften the liver and restore fluid balance. The coordinated use of these three methods， treating both ti and yong， facilitates the separation of dryness and dampness， providing a novel approach to syndrome differentiation and treatment for this subtype of SS.

OBJECTIVE To study the chemical constituents in the root bark of Schisandra sphenanthera and their cytotoxic activ-ities.METHODS The compounds were isolated and purified by silica,Sephadex LH-20 and semi preparative-HPLC and the chem-ical structures were identified by 1 H-NMR,13 C-NMR and MS data analysis.The cytotoxic activities of the compounds were deter-mined by MTT method.RESULTS Twenty lignans were isolated and deduced as:Matairesinol(1),2-Hydroxy-2-(3′,4′-di-hydroxyphenyl)methyl-3-(3″,4″-dimethoxyphenyl)methyl-gamma-butyrolactone(2),(+)-Nortrachelogenin(3),2-Hydroxy-2-(4′-O-β-D-glucopyranosyl-3′-hydroxyphenyl)methyl-3-(3″,4″-dimethoxyphenyl)methyl-γ-butyrolactone(4),Nortracheloside(5),Burselignan(6),(+)-Cycloolivil(7),5-Methoxy-(+)-isolariciresinol(8),(-)-Isolariciresinol 3α-O-β-D-glucopyranoside(9),(+)-9-O-β-D-Glucopyranosyl lyoniresinol(10),(-)-Secoisolariciresinol(11),Licarin A(12),Cedrusin(13),Mataires-inol 4′-O-β-D-glucopyranoside(14),Pregomisin(15),Meso-dihydroguaiaretic acid(16),7S,8R-Erythro-4,9,9′-trihydroxy-3,3′-dimethoxy-8-O-4′-neolignan-7-O-β-D-glucopyranoside(17),Gomisin M2(18),Gomisin M3(19),Pinoresinol(20).Com-pounds 1-3,12,15,16,18 and 19 showed cytotoxic activity against A549,HCT116 and SW620 cell lines with IC50 values ranging from 1.4 to 22.9 μmol·L-1.CONCLUSION Compounds 1-4,6-12,14,17-19 are isolated from the plant for the first time,com-pounds 1-3,12,15,16,18 and 19 exhibit cytotoxic activities.

OBJECTIVE To study the chemical constituents in the root bark of Schisandra sphenanthera and their cytotoxic activ-ities.METHODS The compounds were isolated and purified by silica,Sephadex LH-20 and semi preparative-HPLC and the chem-ical structures were identified by 1 H-NMR,13 C-NMR and MS data analysis.The cytotoxic activities of the compounds were deter-mined by MTT method.RESULTS Twenty lignans were isolated and deduced as:Matairesinol(1),2-Hydroxy-2-(3′,4′-di-hydroxyphenyl)methyl-3-(3″,4″-dimethoxyphenyl)methyl-gamma-butyrolactone(2),(+)-Nortrachelogenin(3),2-Hydroxy-2-(4′-O-β-D-glucopyranosyl-3′-hydroxyphenyl)methyl-3-(3″,4″-dimethoxyphenyl)methyl-γ-butyrolactone(4),Nortracheloside(5),Burselignan(6),(+)-Cycloolivil(7),5-Methoxy-(+)-isolariciresinol(8),(-)-Isolariciresinol 3α-O-β-D-glucopyranoside(9),(+)-9-O-β-D-Glucopyranosyl lyoniresinol(10),(-)-Secoisolariciresinol(11),Licarin A(12),Cedrusin(13),Mataires-inol 4′-O-β-D-glucopyranoside(14),Pregomisin(15),Meso-dihydroguaiaretic acid(16),7S,8R-Erythro-4,9,9′-trihydroxy-3,3′-dimethoxy-8-O-4′-neolignan-7-O-β-D-glucopyranoside(17),Gomisin M2(18),Gomisin M3(19),Pinoresinol(20).Com-pounds 1-3,12,15,16,18 and 19 showed cytotoxic activity against A549,HCT116 and SW620 cell lines with IC50 values ranging from 1.4 to 22.9 μmol·L-1.CONCLUSION Compounds 1-4,6-12,14,17-19 are isolated from the plant for the first time,com-pounds 1-3,12,15,16,18 and 19 exhibit cytotoxic activities.