OBJECTIVE To explore and predict the quality markers （Q-markers） of Compound xiebai capsules for the treatment of chronic obstructive pulmonary disease （COPD） by constituents analysis combined with network pharmacology and molecular docking studies， and to establish the quality standard of Compound xiebai capsules. METHODS UHPLC-TOF-MS was used for qualitative analysis of Compound xiebai capsules， and the candidate Q-markers of Compound xiebai capsules were screened by combining network pharmacology and molecular docking technology. Further， HPLC was applied to establish the fingerprints of 15 batches of Compound xiebai capsules and to conduct quantitative analysis of the main components. RESULTS A total of 51 components were identified from Compound xiebai capsules. Among them， 15 components， namely oxyberberine， methylworenine， coptisine， tetrahydroberberine， epiberberine， berberine， magnoflorine， gandensin， cucurbitacin D， hydroxygenkwan， jatrorrhizine， columbamine， quercetin， cucurbitacin R， and palmatine， were determined as the candidate Q-markers for Compound xiebai capsules in the treatment of COPD. A total of 13 common peaks were calibrated in the fingerprints of 15 batches of Compound xiebai capsules for COPD treatment， with similarity values ranging from 0.976 to 0.999 compared to the reference fingerprint. Seven components were identified among these peaks， namely peak 5 （magnoflorine）， peak 8 （jatrorrhizine）， peak 9 （epiberberine）， peak 10 （columbamine）， peak 11 （coptisine）， peak 12 （palmatine）， and peak 13 （berberine）. Their respective contents were （0.267±0.048）， （0.453±0.084）， （0.572±0.160）， （0.392±0.074）， （1.076±0.273）， （1.477±0.271）， and （6.664±1.249） mg/g （ n ＝3）. CONCLUSIONS This study predicted 15 candidate Q-markers of Compound xiebai capsules in the treatment of COPD and established the fingerprint along with a quantitative determination method for seven major components.

Objective·To determine hyaluronan(HA)expression in the endocrine-resistant microenvironment of estrogen receptor-positive(ER+)breast cancer and elucidate its impact on the acquired resistance.Methods·Chemiluminescent immunoassay was used to quantify HA levels in the culture supernatants of fulvestrant-resistant breast cancer cells.An immunofluorescence(IF)assay was performed to visualize the colocalization of CD44 and HA in MCF7/FulR cells.Using an established adaptive endocrine-resistant breast cancer mouse model,HA expression in resistant breast cancer tissues was assessed by immunohistochemistry(IHC)assay.Single-cell RNA sequencing(scRNA-seq)and RNA sequencing(RNA-seq)were conducted to examine transcriptomic profiles and alterations in HA-related genes in resistant breast cancer cells.Flow cytometry(FCM)was utilized to measure the proportion of CD44+CD24-cells in MCF7/FulR.The correlation between HA synthesis genes and cell stemness was investigated in clinical ER+breast cancers from GEO data sets.Hyaluronidase(HAase)treatment was applied to remove the"HA coat",and RT-qPCR and Western blotting analysis were carried out to monitor changes in stemness-related molecules.CCK-8 assays,flow cytometry(FCM),and Hoechst 33258 staining were performed to determine changes in apoptosis and fulvestrant efficiency after HAase treatment.Results·IF results revealed that compared with MCF7 cells,the"HA coat"on the surface of MCF7/FulR cells was significantly thickened.IHC demonstrated markedly increased HA retention in fulvestrant-resistant mouse breast cancer tissues.ScRNA-seq and RNA-seq analyses indicated elevated expression of stemness-related genes and HA synthesis-associated genes in fulvestrant-resistant breast cancer cells.Correlation analysis revealed a positive association between HA synthesis and cancer stemness in ER+breast cancer.IF and RT-qPCR results demonstrated that removing the HA coating from the surface of MCF7/FulR cells led to a significant reduction in the expression of stemness-related molecules;concurrently,CCK-8 assays,FCM analysis,and Hoechst 33258 staining revealed that"HA coat"clearance reduced MCF7/FulR'tolerance to fulvestrant and increased apoptosis.Conclusion·Endocrine-resistant breast cancer cells develop an enriched"HA coat",which promotes stemness in fulvestrant-resistant tumors.Disruption of this HA coat through HAase treatment effectively reduces cell stemness,induces apoptosis,and re-sensitizes breast cancer cells to fulvestrant.

Objective:To explore the clinical characteristics and prognostic factors of primary bladder adenocarcinoma（ACB）.Methods:The clinical data of 33 patients with primary ACB who underwent surgical treatment in the Department of Urology of Peking University Third Hospital from July 2003 to January 2024 were retrospectively analyzed. There were 5 females and 28 males，with an average age of（61.3 ± 11.5）years. Twelve patients had comorbidities（6 hypertension，2 coronary heart disease，5 diabetes mellitus，and 3 cerebrovascular diseases）and 12 had a smoking history. The mean body mass index（BMI）was（24.8 ± 3.2）kg/m 2. The maximum tumor diameter measured by enhanced computed tomography（CT）was（29.7 ± 12.7）mm. The preoperative neutrophil-to-lymphocyte ratio（NLR）was 3.4 ± 3.2，and the systemic immune-inflammation index（SII）was（582 ± 496）× 10 9/L.Patients were divided into two groups according to the surgical approach：the radical cystectomy group（ n = 23）and the bladder-sparing group（ n = 10）. For the radical cystectomy group，there were 19 males and 4 females，with a mean age of（59.9 ± 12.6）years. Five patients had comorbidities（3 hypertension，1 coronary heart disease，1 diabetes mellitus，and 2 cerebrovascular diseases）. Eight patients had a smoking history，with a mean BMI of（25.2 ± 3.5）kg/m 2. The maximum tumor diameter was（33.6 ± 10.9）mm，the preoperative NLR was 3.5 ± 3.5，and the SII was（618 ± 558）× 10 9/L. For the bladder-sparing group，there were 9 males and 1 female，with a mean age of（64.5 ± 8.2）years. Seven patients had comorbidities（3 hypertension，1 coronary heart disease，4 diabetes mellitus，and 1 cerebrovascular diseases）. Four patients had a smoking history，with a mean BMI of（23.5 ± 2.3）kg/m 2. The maximum tumor diameter was（20.7 ± 12.5）mm，the preoperative NLR was 3.1 ± 2.2，and the SII was（501 ± 323）× 10 9/L. Statistically significant differences were observed between the two groups in terms of comorbidities（ P = 0.008）and maximum tumor diameter（ P = 0.006），while no significant differences were found in other data（ P > 0.05）. The Kaplan-Meier survival curve was drawn，and Cox regression was used to analyze the prognostic factors of progression-free survival（PFS）and overall survival（OS）of patients. Results:Among the 33 patients，low-grade adenocarcinoma and high-grade adenocarcinoma accounted for 60.6% and 39.4% respectively according to the postoperative pathology，and 3 patients had positive surgical margins. There were 22 cases of muscle-invasive bladder adenocarcinoma，5 cases of lymph node metastasis，and 1 case of distant metastasis. The patients in tumor stages Ⅰ—Ⅳ were 9 cases（27.3%），8 cases（24.2%），7 cases（21.2%），and 9 cases（27.3%）respectively. Nine patients received postoperative adjuvant therapy，including 6 with adjuvant chemotherapy，2 with adjuvant chemotherapy combined with radiotherapy，and 1 with adjuvant immunotherapy. In the radical cystectomy group（ n = 23），there were 13 cases of low-grade and 10 cases of high-grade pathological grading，2 cases with positive margins，19 cases of muscle-invasive bladder adenocarcinoma，5 cases of lymph node metastasis，1 case of distant metastasis，and 5 patients received adjuvant therapy（4 cases of adjuvant chemotherapy，and 1 case of adjuvant immunotherapy）. In the bladder-sparing group（ n = 10），there were 7 cases of low-grade，3 cases of high-grade pathological grading，1 case with positive margins，3 cases of muscle-invasive bladder adenocarcinoma，zero lymph node or distant metastasis，and 4 patients received adjuvant therapy（2 cases of adjuvant chemotherapy，and 2 cases of combined adjuvant chemotherapy and radiotherapy）. A statistically significant difference was found in the proportion of muscle-invasive bladder adenocarcinoma between the two groups（ P = 0.006），while no significant differences were observed in other data（ P > 0.05）.The median follow-up duration of the patients was 28.0 months，the median PFS was 86.0 months，and the median OS was 90.0 months. The 2-year PFS and OS were 65.4% and 73.1% respectively. The 5-year PFS and OS were 54.2% and 56.5% respectively. The Kaplan-Meier survival analysis showed that there were no significant differences in PFS（ P = 0.777）and OS（ P = 0.585）between the radical cystectomy group and the bladder-preserving group. Female（ P = 0.011），BMI < 25 kg/m2（ P = 0.038），and positive surgical margins（ P < 0.01）were associated with poorer PFS. Aged ≥ 70 years（ P = 0.003），lymph node metastasis（ P = 0.041），and positive surgical margins（ P = 0.025）were associated with poorer OS，and patients in the adjuvant therapy group had better OS（ P = 0.005）. Multivariate Cox regression analysis indicated that positive surgical margins（HR 10.2， P = 0.012）were an independent impact factor for PFS，and positive surgical margins（HR 39.8， P = 0.001）and adjuvant therapy（HR 0.12， P = 0.021）were independent impact factors for OS. Conclusions:Positive surgical margins and adjuvant therapy are independent impact factors for the prognosis of patients with primary ACB.

Objective·To determine hyaluronan(HA)expression in the endocrine-resistant microenvironment of estrogen receptor-positive(ER+)breast cancer and elucidate its impact on the acquired resistance.Methods·Chemiluminescent immunoassay was used to quantify HA levels in the culture supernatants of fulvestrant-resistant breast cancer cells.An immunofluorescence(IF)assay was performed to visualize the colocalization of CD44 and HA in MCF7/FulR cells.Using an established adaptive endocrine-resistant breast cancer mouse model,HA expression in resistant breast cancer tissues was assessed by immunohistochemistry(IHC)assay.Single-cell RNA sequencing(scRNA-seq)and RNA sequencing(RNA-seq)were conducted to examine transcriptomic profiles and alterations in HA-related genes in resistant breast cancer cells.Flow cytometry(FCM)was utilized to measure the proportion of CD44+CD24-cells in MCF7/FulR.The correlation between HA synthesis genes and cell stemness was investigated in clinical ER+breast cancers from GEO data sets.Hyaluronidase(HAase)treatment was applied to remove the"HA coat",and RT-qPCR and Western blotting analysis were carried out to monitor changes in stemness-related molecules.CCK-8 assays,flow cytometry(FCM),and Hoechst 33258 staining were performed to determine changes in apoptosis and fulvestrant efficiency after HAase treatment.Results·IF results revealed that compared with MCF7 cells,the"HA coat"on the surface of MCF7/FulR cells was significantly thickened.IHC demonstrated markedly increased HA retention in fulvestrant-resistant mouse breast cancer tissues.ScRNA-seq and RNA-seq analyses indicated elevated expression of stemness-related genes and HA synthesis-associated genes in fulvestrant-resistant breast cancer cells.Correlation analysis revealed a positive association between HA synthesis and cancer stemness in ER+breast cancer.IF and RT-qPCR results demonstrated that removing the HA coating from the surface of MCF7/FulR cells led to a significant reduction in the expression of stemness-related molecules;concurrently,CCK-8 assays,FCM analysis,and Hoechst 33258 staining revealed that"HA coat"clearance reduced MCF7/FulR'tolerance to fulvestrant and increased apoptosis.Conclusion·Endocrine-resistant breast cancer cells develop an enriched"HA coat",which promotes stemness in fulvestrant-resistant tumors.Disruption of this HA coat through HAase treatment effectively reduces cell stemness,induces apoptosis,and re-sensitizes breast cancer cells to fulvestrant.

Objective To explore the potential impact of unhealthy diets on cardiovascular diseases and all-cause mortality.Methods This study included the individuals aged 35-70 years from 45 cities and 70 rural communities across 12 provinces in China,as part of the Prospective Urban Rural Epidemiology(PURE)study.Dietary habits were assessed using a food frequency questionnaire.The dietary health status was scored using the Alternative Healthy Eating Index(AHEI),with participants in the lowest tertile of AHEI being categorized into the unhealthy diet group,while others were classified as the healthy diet group.The primary endpoints included major cardiovas-cular diseases(myocardial infarction,stroke,or heart failure)and all-cause mortality.Cox proportional hazard models were used to estimate hazard ratios(HR)for each group.Results A total of 40 925 participants were in-cluded in the study,with a median follow-up time of 11.9 years(interquartile range 9.6-12.6 years).During this period,2 066 deaths and 3 099 cases of major cardiovascular diseases were reported.The results showed that un-healthy diet increased the risk of major cardiovascular diseases by 10%(HR=1.10,95%CI:1.02-1.20,P＜0.05)and all-cause mortality by 7%(HR=1.07,95%CI:1.00-1.18,P＜0.05).Among male residents,un-healthy diet did not increase the risk of major cardiovascular diseases or all-cause mortality.However,among female residents,those with an unhealthy diet had a higher risk of major cardiovascular diseases(HR=1.12,95%CI:1.00-1.25,P＜0.05)and all-cause mortality(HR=1.26,95%CI:1.08-1.46,P＜0.05)compared to those with a healthy diet.Conclusions Unhealthy diet increases the risk of major cardiovascular diseases and all-cause mortality,particularly among women.There is a need to raise awareness about healthy dietary to prevent death and the occurrence of major cardiovascular diseases.

[Objective] To explore the influencing factors of quality monitoring index on the nucleic acid pooling detection mode and continuously improve the detection quality of nucleic acid laboratory. [Methods] The quality monitoring indicators (NAT reactive rate, NAT resolution reactive rate, NAT invalid batch rate, NAT invalid result rate, equipment failure rate) and causes of invalidity in our laboratory from January 1, 2020 to December 31, 2022 were retrospectively analyzed. The quality monitoring indicators of the laboratory during 2020 to 2022 were compared longitudinally. The quality monitoring indicators of the laboratory in 2022 were compared horizontally with the overall level in Shandong for the same period to find the differences. [Results] From 2020 to 2022, a total of 218 686 samples were detected, the NAT reactive rate was 0.15‰ (32 samples in total), the resolution reactive rate was 39.02%, the invalid batch rate was 1.06%, the invalid result rate was 1.18%, and the equipment failure rate was 3.58%. There were no differences in the NAT reactive rate, NAT resolution reactive rate and NAT invalid batch rate among different years (P>0.05), but there were differences in the invalid result rate (P<0.05). Equipment failure was the main cause of invalid results (56.53%). Compared with other laboratories in Shandong, there were differences in the NAT reactive rate and invalid result rate (P<0.05). There were differences in the reaction rate, resolution rate and invalid result rate among different reagents (P<0.05). Compared with other two laboratories using the same manufacturer's reagent, there were differences in the reactive rate and invalid result rate (P<0.05), but no difference in the resolution rate and invalid batch rate (P>0.05). [Conclusion] Establishing quality indexes for process control and regular analysis can timely detect potential risks in laboratory operation. The use of quality indicators to implement self-comparison and inter-laboratory comparison can help the laboratory systematically and scientifically evaluate its own operating status and formulate corresponding quality management strategies, thereby improving the laboratory's testing capacity and ensure the safety of blood use.

BACKGROUND:We have successfully established an animal model of small ear pig in southern Yunnan province with internal tension relieving technique combined with autologous Achilles tendon for anterior cruciate ligament reconstruction,and verified the stability and reliability of the model.However,whether internal tension relieving technique can promote the ligamentalization process of autologous Achilles tendon graft has not been studied. OBJECTIVE:To investigate the differences in the process of ligamentalization between conventional reconstruction and internal reduction reconstruction of the anterior cruciate ligament by gross view,histology and electron microscopy. METHODS:Thirty adult female small ear pigs in southern Yunnan province were selected.Anterior cruciate ligament reconstruction was performed on the left knee joint with the ipsilateral knee Achilles tendon(n=30 in the normal group),and anterior cruciate ligament reconstruction was performed on the right knee joint with the ipsilateral knee Achilles tendon combined with the internal relaxation and enhancement system(n=30 in the relaxation group).The autogenous right forelimb was used as the control group;the anterior cruciate ligament was exposed but not severed or surgically treated.At 12,24,and 48 weeks after surgery,10 animals were sacrificed,respectively.The left and right knee joint specimens were taken for gross morphological observation to evaluate the graft morphology.MAS score was used to evaluate the excellent and good rate of the ligament at each time point.Hematoxylin-eosin staining was used to evaluate the degree of ligament graft vascularization.Collagen fibers and nuclear morphology were observed,and nuclear morphology was scored.Ultrastructural remodeling was evaluated by scanning electron microscopy and transmission electron microscopy. RESULTS AND CONCLUSION:(1)The ligament healing shape of the relaxation group was better at various time points after surgery,and the excellent and good rate of MAS score was higher(P<0.05).Moreover,the relaxation group could obtain higher ligament vascularization score(P<0.05).(2)The arrangement of collagen bundles and fiber bundles in the two groups gradually tended to be orderly,and the transverse fiber connections between collagen gradually increased and thickened,suggesting that the strength and shape degree of the grafts were gradually improved,but the ligament remodeling in the relaxation group was always faster than that in the normal group at various time points after surgery.(3)The diameter,distribution density,and arrangement degree of collagen fibers in the relaxation group were better than those in the normal group at all time points,especially in the comparison of collagen fiber diameter between and within the relaxation group(P<0.05).

Objective:To observe the improving effect of acupuncture and Tuina(Chinese therapeutic massage)combined with Ba Duan Jin(Eight-brocade Exercise)on motor function in patients with Parkinson disease(PD)in the early and middle stages.Methods:Seventy patients with early-or middle-stage PD were randomly divided into a control group and an observation group,with 35 patients in each group.The control group was given conventional treatment such as oral levodopa hydrochloride tablets;the observation group was treated with acupuncture,Tuina,and Ba Duan Jin in addition to the control group's treatment.Acupuncture and Tuina were administered twice a week,and Ba Duan Jin was performed five times a week.Each course of treatment lasted 4 weeks,and a total of 3 courses were completed.The changes in the unified Parkinson disease rating scale Ⅲ(UPDRS Ⅲ)score,Berg balance scale(BBS)score,15-meter walking speed,and 6-minute walk distance were compared between the two groups.Results:The total effective rate in the observation group was 93.6%,which was significantly higher than 45.7%in the control group(P<0.01).After 4,8,and 12 weeks of treatment,the UPDRS Ⅲ score in the observation group was significantly lower than the previous score(P<0.05);the BBS score was significantly higher than the previous score(P<0.05),and the 6-minute walk distance also significantly increased compared to the previous result(P<0.05).After 8 and 12 weeks of treatment,the 15-meter walking speed in the observation group was significantly higher than that before treatment and the previous measurement(P<0.05).In comparison with the control group,the observation group showed significantly lower UPDRS Ⅲ scores after 8 and 12 weeks of treatment(P<0.05).Additionally,after 4,8,and 12 weeks of treatment,the observation group had significantly higher BBS scores,15-meter walking speed,and the 6-minute walk distance than the control group(P<0.05).There were no significant differences in the above indicators before and after treatment in the control group(P>0.05).Conclusion:The addition of acupuncture,Tuina,and Ba Duan Jin to Western medication treatment can effectively improve motor function,balance ability,and walking ability in patients with early-or middle-stage PD.

Objective:To explore the clinical characteristics and prognostic factors of primary bladder adenocarcinoma（ACB）.Methods:The clinical data of 33 patients with primary ACB who underwent surgical treatment in the Department of Urology of Peking University Third Hospital from July 2003 to January 2024 were retrospectively analyzed. There were 5 females and 28 males，with an average age of（61.3 ± 11.5）years. Twelve patients had comorbidities（6 hypertension，2 coronary heart disease，5 diabetes mellitus，and 3 cerebrovascular diseases）and 12 had a smoking history. The mean body mass index（BMI）was（24.8 ± 3.2）kg/m 2. The maximum tumor diameter measured by enhanced computed tomography（CT）was（29.7 ± 12.7）mm. The preoperative neutrophil-to-lymphocyte ratio（NLR）was 3.4 ± 3.2，and the systemic immune-inflammation index（SII）was（582 ± 496）× 10 9/L.Patients were divided into two groups according to the surgical approach：the radical cystectomy group（ n = 23）and the bladder-sparing group（ n = 10）. For the radical cystectomy group，there were 19 males and 4 females，with a mean age of（59.9 ± 12.6）years. Five patients had comorbidities（3 hypertension，1 coronary heart disease，1 diabetes mellitus，and 2 cerebrovascular diseases）. Eight patients had a smoking history，with a mean BMI of（25.2 ± 3.5）kg/m 2. The maximum tumor diameter was（33.6 ± 10.9）mm，the preoperative NLR was 3.5 ± 3.5，and the SII was（618 ± 558）× 10 9/L. For the bladder-sparing group，there were 9 males and 1 female，with a mean age of（64.5 ± 8.2）years. Seven patients had comorbidities（3 hypertension，1 coronary heart disease，4 diabetes mellitus，and 1 cerebrovascular diseases）. Four patients had a smoking history，with a mean BMI of（23.5 ± 2.3）kg/m 2. The maximum tumor diameter was（20.7 ± 12.5）mm，the preoperative NLR was 3.1 ± 2.2，and the SII was（501 ± 323）× 10 9/L. Statistically significant differences were observed between the two groups in terms of comorbidities（ P = 0.008）and maximum tumor diameter（ P = 0.006），while no significant differences were found in other data（ P > 0.05）. The Kaplan-Meier survival curve was drawn，and Cox regression was used to analyze the prognostic factors of progression-free survival（PFS）and overall survival（OS）of patients. Results:Among the 33 patients，low-grade adenocarcinoma and high-grade adenocarcinoma accounted for 60.6% and 39.4% respectively according to the postoperative pathology，and 3 patients had positive surgical margins. There were 22 cases of muscle-invasive bladder adenocarcinoma，5 cases of lymph node metastasis，and 1 case of distant metastasis. The patients in tumor stages Ⅰ—Ⅳ were 9 cases（27.3%），8 cases（24.2%），7 cases（21.2%），and 9 cases（27.3%）respectively. Nine patients received postoperative adjuvant therapy，including 6 with adjuvant chemotherapy，2 with adjuvant chemotherapy combined with radiotherapy，and 1 with adjuvant immunotherapy. In the radical cystectomy group（ n = 23），there were 13 cases of low-grade and 10 cases of high-grade pathological grading，2 cases with positive margins，19 cases of muscle-invasive bladder adenocarcinoma，5 cases of lymph node metastasis，1 case of distant metastasis，and 5 patients received adjuvant therapy（4 cases of adjuvant chemotherapy，and 1 case of adjuvant immunotherapy）. In the bladder-sparing group（ n = 10），there were 7 cases of low-grade，3 cases of high-grade pathological grading，1 case with positive margins，3 cases of muscle-invasive bladder adenocarcinoma，zero lymph node or distant metastasis，and 4 patients received adjuvant therapy（2 cases of adjuvant chemotherapy，and 2 cases of combined adjuvant chemotherapy and radiotherapy）. A statistically significant difference was found in the proportion of muscle-invasive bladder adenocarcinoma between the two groups（ P = 0.006），while no significant differences were observed in other data（ P > 0.05）.The median follow-up duration of the patients was 28.0 months，the median PFS was 86.0 months，and the median OS was 90.0 months. The 2-year PFS and OS were 65.4% and 73.1% respectively. The 5-year PFS and OS were 54.2% and 56.5% respectively. The Kaplan-Meier survival analysis showed that there were no significant differences in PFS（ P = 0.777）and OS（ P = 0.585）between the radical cystectomy group and the bladder-preserving group. Female（ P = 0.011），BMI < 25 kg/m2（ P = 0.038），and positive surgical margins（ P < 0.01）were associated with poorer PFS. Aged ≥ 70 years（ P = 0.003），lymph node metastasis（ P = 0.041），and positive surgical margins（ P = 0.025）were associated with poorer OS，and patients in the adjuvant therapy group had better OS（ P = 0.005）. Multivariate Cox regression analysis indicated that positive surgical margins（HR 10.2， P = 0.012）were an independent impact factor for PFS，and positive surgical margins（HR 39.8， P = 0.001）and adjuvant therapy（HR 0.12， P = 0.021）were independent impact factors for OS. Conclusions:Positive surgical margins and adjuvant therapy are independent impact factors for the prognosis of patients with primary ACB.

Objective To investigate the effects of cucurbitacin B (CucB) on alleviating skin lesions and inflammation in psoriasis mice via the cGAS-STING signaling pathway. Methods The expression of genes associated with the cGAS-STING signaling pathway in psoriatic lesions and non-lesional skin was analyzed, and hallmark gene set enrichment analysis was performed. The cytotoxicity of CucB on BMDMs was evaluated using the CCK-8 assay. The expression levels of genes and proteins related to the cGAS-STING signaling pathway, along with the secretion of inflammatory cytokines, were measured at different concentrations of CucB using quantitative PCR, Western blotting, and ELISA. Imiquimod-induced psoriasis BALB/c mice were divided into four groups: normal group, model group, low-dose CucB group [0.1 mg/ (kg.d)], and high-dose CucB group [0.4 mg/ (kg.d)], with five mice per group. PASI scoring was performed to assess the severity of psoriasis after 6 days of treatment, and HE staining was conducted to observe pathological damage. Meanwhile, the mRNA levels of inflammatory cytokines and their secretion were detected by qPCR and ELISA. Results Most cGAS-STING signaling-related genes were upregulated in lesional skin of psoriasis patients, and the hallmark gene set enrichment analysis revealed that the most significantly upregulated genes were primarily associated with immune response signaling pathways. CucB inhibited dsDNA-induced phosphorylation of interferon regulatory factor 3 (IRF3) and STING proteins in both bone-marrow derived macrophages(BMDMs) and THP-1 cells. CucB also suppressed dsDNA-induced mRNA expression of IFNB1, TNF, IFIT1, CXCL10, ISG15, and reduced the secretion of cytokines such as IFN-β, IL-1β, and TNF-α in THP-1 cells. In the imiquimod-induced psoriasis mouse model, CucB treatment reduced psoriatic symptoms, alleviated skin lesions, and attenuated inflammation. ELISA and qPCR results showed that CucB significantly reduced serum secretion levels of IL-6, TNF-α, and IL-1β, as well as the mRNA levels of IL23A, IL1B, IL6, TNF, and IFNB1. Conclusion CucB inhibits cytoplasmic DNA-induced activationc of the GAS-STING pathway. CucB significantly attenuates skin lesions and inflammation in IMQ-induced psoriatic mice, and the potential molecular mechanism may be related to the down-regulation of the cGAS-STING pathway.
Animals ; Psoriasis/pathology* ; Signal Transduction/drug effects* ; Membrane Proteins/genetics* ; Mice ; Nucleotidyltransferases/genetics* ; Disease Models, Animal ; Mice, Inbred BALB C ; Skin/metabolism* ; Triterpenes/therapeutic use* ; Humans ; Cytokines/metabolism* ; Inflammation/drug therapy* ; Male