1.Total flavonoids of Pterocarya hupehensis Skan alleviate DSS-induced ul-cerative colitis in mice by modulating macrophage polarization
Guoqing CHEN ; Xiaorong LIU ; Jin JIN ; Dong YAN ; Renjia LIU ; Shan XIANG ; Lin YUAN ; Yang XIANG ; Hao WU ; Xiulan SHEN
Chinese Journal of Pathophysiology 2025;41(6):1181-1189
AIM:To investigate the effects of total flavonoids of Pterocarya hupehensis Skan(PHSTF)on dex-tran sulfate sodium(DSS)-induced ulcerative colitis(UC)mouse model and lipopolysaccharide(LPS)-stimulated RAW264.7 macrophages.METHODS:Thirty-six male C57BL/6J mice(6 to 8 weeks old,SPF grade)were randomly di-vided into 6 groups:negative control(NC)group,3%DSS-induced model group,mesalazine(300 mg·kg-1·d-1)group,and low-dose(62.5 mg·kg-1·d-1),medium-dose(125 mg·kg-1·d-1)and high-dose(250 mg·kg-1·d-1)PHSTF treatment groups,with 6 mice in each group.The mice in NC group received distilled water,while those in other groups were treated with a 3%DSS solution for 7 d to induce the UC model.On the 1st day of DSS administration,the mice in treatment groups received the corresponding agents via oral gavage for 10 d,while those in NC and model groups were gavaged with distilled water.Throughout the study,the effects of PHSTF on body weight,fecal blood,and colon length were measured and recorded daily.Histopathological changes in colon tissues were assessed using hematoxylin-eosin staining.The levels of the pro-inflammatory cytokine interleukin-1β(IL-1β)and the anti-inflammatory cytokine IL-10 in colon tissues were quantified using ELISA.The LPS-induced RAW264.7 macrophage model was employed to evaluate the cellular effects of PHSTF.Cell viability was assessed by CCK-8 assay,and cell morphology was observed under a microscope.The mRNA expression of inflammatory markers[IL-1β,inducible nitric oxide synthase(iNOS),IL-10 and arginase-1(Arg-1)]was measured by RT-qPCR.Western blot and immunofluorescence double labeling were used to detect the protein expression of macrophage polarization markers(iNOS,CD206 and Arg-1).Finally,immunohistochemistry(IHC)was utilized to as-sess protein expression of iNOS in colon tissues.RESULTS:Compared to the DSS-induced UC model group,PHSTF sig-nificantly improved several parameters,including weight loss(P<0.05),rectal bleeding,and colon shortening in DSS-treated mice.PHSTF also reduced histopathological damage and inflammatory cell infiltration in the colon.It decreased IL-1β levels(P<0.05)and increased IL-10 levels(P<0.05)in colon tissues.In LPS-induced RAW264.7 cells,PHSTF reduced the mRNA expression of IL-1β and iNOS(P<0.01),while upregulating the mRNA expression of IL-10 and Arg-1(P<0.01).Additionally,PHSTF decreased iNOS protein expression(P<0.01)and elevated the expression of Arg-1 and CD206 proteins(P<0.01).IHC analysis further confirmed that PHSTF downregulated iNOS protein expression in colon tissues.CONCLUSION:Treatment with PHSTF promotes the polarization of macrophages from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype,thereby alleviating inflammation in colon tissue and ameliorating ulcer-ative colitis in mice.
2.Analysis of liver histological characteristics and clinically related factors in patients with inactive HBsAg carriers
Xinyang ZHANG ; Shan REN ; Sujun ZHENG ; Rongshan FAN ; Qingfa RUAN ; Wenqi HUANG ; Haibing GAO ; Yao XIE ; Minghui LI ; Xiulan XUE ; Fang YANG ; Junliang FU ; Xinyue CHEN
Chinese Journal of Hepatology 2025;33(7):660-666
Objective:To analyze the liver histological characteristics and clinically related factors in inactive hepatitis B surface antigen (HBsAg) carriers (IHC), and also explore whether antiviral treatment is necessary for IHC, as defined in the 2022 version of the hepatitis B prevention and treatment guidelines.Methods:A multicenter, retrospective cohort study was conducted. Two hundred and thirty-one IHC cases who underwent liver biopsy histopathological examination in nine medical institutions, including Beijing Youan Hospital affiliated with Capital Medical University, from January 2018 to December 2023 were included. General informative data, clinical serological markers, and transient elastography (TE) examination results were collected. Patients were divided into a positive (148 cases) and a negative group (83 cases) according to the results of hepatitis B virus (HBV) DNA detection. The differences in liver pathological inflammatory activity (G) and liver fibrosis stage (S) were analyzed between the two groups to explore the correlation between liver tissue conditions and clinically related factors. Comparsions of normally distributed continwous data, skeukd continuous data, and categorical data between groups are performed using t tests, Mann-Whitney U tests and χ2 tests, respectively. Results:The age of 231 IHC cases was 43 (38, 51) years old, with 95.2% (220/231) aged ≥30 years, and males accounted for 64.9% (150/231). HBsAg and HBV DNA levels were 131.9 (20.8, 400.9) IU/mL and 94.0 (0, 448.5) IU/mL, respectively, of which 35.9% (83/231) were HBV DNA negative (<20 IU/mL). The remarkable proportions of G≥2, S≥2, and liver injury (G≥2 and/or S≥2) in liver tissue were 16.5% (38/231), 29% (67/231), and 35.9% (83/231), respectively. The S≥2 proportion was significantly higher in the HBV DNA-negative group than the positive group (42.2% vs. 21.6%, P<0.001), and it mainly occurred in the population cohort over 30 years old (44.9% vs. 31.0%, P=0.04). The liver stiffness measurement (LSM), aspartate transaminase to platelet ratio index (APRI), and platelet (PLT) were significantly higher in the S≥2 group than the S<2 group ( P<0.05). Conclusion:Clinicians can comprehensively evaluate the degree of liver fibrosis in IHC based on clinical factors such as age, PLT, APRI, and LSM, even if the liver histological results are lacking. The China 2022 version guidelines define that nearly half of IHC has histological indications for antiviral therapy, and liver biopsy and prompt treatment can be recommended.
3.Additional benefits of pelvic floor proprioceptive training combined with conventional therapy in the treatment of female stress urinary incontinence.
Xiulan ZHANG ; Liping ZHU ; Xiaoling ZENG ; Zhaoxue LIU ; Shuo YANG ; Hong ZHANG ; Wenguang YAN ; Xuhong LI
Journal of Central South University(Medical Sciences) 2025;50(8):1385-1397
OBJECTIVES:
Stress urinary incontinence (SUI) is a common condition among women that severely impairs quality of life. Pelvic floor proprioceptive training (PFPT) has attracted increasing attention for its potential to enhance pelvic floor muscle function and alleviate SUI symptoms. This study aims to observe and compare the clinical efficacy of PFPT combined with electroacupuncture, electrical stimulation, and biofeedback therapy versus conventional therapy consisting of electroacupuncture, electrical stimulation, and biofeedback alone in women with SUI, and to explore the role of PFPT in improving symptom and functional outcomes.
METHODS:
In this randomized controlled trial, 72 women with mild to moderate SUI were recruited from the Department of Rehabilitation Medicine at Third Xiangya Hospital, Central South University, between December 2021 and October 2023. Participants were randomly assigned to an experimental group (n=36) or a control group (n=36). Both groups received health education. The control group underwent electroacupuncture combined with electrical stimulation and biofeedback therapy, while the experimental group additionally received PFPT 3 times per week for 4 weeks. The primary outcome was assessed using the International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF). Secondary outcomes included pelvic floor muscle strength, bladder neck mobility, and balance ability. The ICIQ-SF was reassessed at 1, 3, 6, and 12 months post-treatment.
RESULTS:
Both groups showed statistically significant improvements in all parameters after treatment (all P<0.05). However, there were no statistically significant differences between groups in most measures (all P>0.05). The experimental group demonstrated longer single-leg stance duration with eyes closed than the control group (left leg: P=0.026; right leg: P=0.006), with a significant increase from baseline (P<0.001). At 6 months post-treatment, the cure rate in the experimental group was significantly higher than that in the control group (P=0.037).
CONCLUSIONS
Conventional therapy effectively improves SUI symptoms, but adding PFPT provides notable additional benefits, including enhanced balance ability and sustained mid-term cure rates. These findings suggest that PFPT is a valuable adjunct to standard SUI management strategies.
Humans
;
Female
;
Urinary Incontinence, Stress/physiopathology*
;
Pelvic Floor/physiopathology*
;
Middle Aged
;
Biofeedback, Psychology
;
Adult
;
Exercise Therapy/methods*
;
Proprioception
;
Electroacupuncture/methods*
;
Quality of Life
;
Electric Stimulation Therapy/methods*
;
Treatment Outcome
;
Combined Modality Therapy
4.Erratum: Author correction to "Sphingosine-1-phosphate, a novel TREM2 ligand, promotes microglial phagocytosis to protect against ischemic brain injury" Acta Pharm Sin B 12 (2022) 1885-1898.
Tengfei XUE ; Juan JI ; Yuqin SUN ; Xinxin HUANG ; Zhenyu CAI ; Jin YANG ; Wei GUO ; Ruobing GUO ; Hong CHENG ; Xiulan SUN
Acta Pharmaceutica Sinica B 2025;15(5):2813-2814
[This corrects the article DOI: 10.1016/j.apsb.2021.10.012.].
5.Total flavonoids of Pterocarya hupehensis Skan alleviate DSS-induced ul-cerative colitis in mice by modulating macrophage polarization
Guoqing CHEN ; Xiaorong LIU ; Jin JIN ; Dong YAN ; Renjia LIU ; Shan XIANG ; Lin YUAN ; Yang XIANG ; Hao WU ; Xiulan SHEN
Chinese Journal of Pathophysiology 2025;41(6):1181-1189
AIM:To investigate the effects of total flavonoids of Pterocarya hupehensis Skan(PHSTF)on dex-tran sulfate sodium(DSS)-induced ulcerative colitis(UC)mouse model and lipopolysaccharide(LPS)-stimulated RAW264.7 macrophages.METHODS:Thirty-six male C57BL/6J mice(6 to 8 weeks old,SPF grade)were randomly di-vided into 6 groups:negative control(NC)group,3%DSS-induced model group,mesalazine(300 mg·kg-1·d-1)group,and low-dose(62.5 mg·kg-1·d-1),medium-dose(125 mg·kg-1·d-1)and high-dose(250 mg·kg-1·d-1)PHSTF treatment groups,with 6 mice in each group.The mice in NC group received distilled water,while those in other groups were treated with a 3%DSS solution for 7 d to induce the UC model.On the 1st day of DSS administration,the mice in treatment groups received the corresponding agents via oral gavage for 10 d,while those in NC and model groups were gavaged with distilled water.Throughout the study,the effects of PHSTF on body weight,fecal blood,and colon length were measured and recorded daily.Histopathological changes in colon tissues were assessed using hematoxylin-eosin staining.The levels of the pro-inflammatory cytokine interleukin-1β(IL-1β)and the anti-inflammatory cytokine IL-10 in colon tissues were quantified using ELISA.The LPS-induced RAW264.7 macrophage model was employed to evaluate the cellular effects of PHSTF.Cell viability was assessed by CCK-8 assay,and cell morphology was observed under a microscope.The mRNA expression of inflammatory markers[IL-1β,inducible nitric oxide synthase(iNOS),IL-10 and arginase-1(Arg-1)]was measured by RT-qPCR.Western blot and immunofluorescence double labeling were used to detect the protein expression of macrophage polarization markers(iNOS,CD206 and Arg-1).Finally,immunohistochemistry(IHC)was utilized to as-sess protein expression of iNOS in colon tissues.RESULTS:Compared to the DSS-induced UC model group,PHSTF sig-nificantly improved several parameters,including weight loss(P<0.05),rectal bleeding,and colon shortening in DSS-treated mice.PHSTF also reduced histopathological damage and inflammatory cell infiltration in the colon.It decreased IL-1β levels(P<0.05)and increased IL-10 levels(P<0.05)in colon tissues.In LPS-induced RAW264.7 cells,PHSTF reduced the mRNA expression of IL-1β and iNOS(P<0.01),while upregulating the mRNA expression of IL-10 and Arg-1(P<0.01).Additionally,PHSTF decreased iNOS protein expression(P<0.01)and elevated the expression of Arg-1 and CD206 proteins(P<0.01).IHC analysis further confirmed that PHSTF downregulated iNOS protein expression in colon tissues.CONCLUSION:Treatment with PHSTF promotes the polarization of macrophages from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype,thereby alleviating inflammation in colon tissue and ameliorating ulcer-ative colitis in mice.
6.Analysis of liver histological characteristics and clinically related factors in patients with inactive HBsAg carriers
Xinyang ZHANG ; Shan REN ; Sujun ZHENG ; Rongshan FAN ; Qingfa RUAN ; Wenqi HUANG ; Haibing GAO ; Yao XIE ; Minghui LI ; Xiulan XUE ; Fang YANG ; Junliang FU ; Xinyue CHEN
Chinese Journal of Hepatology 2025;33(7):660-666
Objective:To analyze the liver histological characteristics and clinically related factors in inactive hepatitis B surface antigen (HBsAg) carriers (IHC), and also explore whether antiviral treatment is necessary for IHC, as defined in the 2022 version of the hepatitis B prevention and treatment guidelines.Methods:A multicenter, retrospective cohort study was conducted. Two hundred and thirty-one IHC cases who underwent liver biopsy histopathological examination in nine medical institutions, including Beijing Youan Hospital affiliated with Capital Medical University, from January 2018 to December 2023 were included. General informative data, clinical serological markers, and transient elastography (TE) examination results were collected. Patients were divided into a positive (148 cases) and a negative group (83 cases) according to the results of hepatitis B virus (HBV) DNA detection. The differences in liver pathological inflammatory activity (G) and liver fibrosis stage (S) were analyzed between the two groups to explore the correlation between liver tissue conditions and clinically related factors. Comparsions of normally distributed continwous data, skeukd continuous data, and categorical data between groups are performed using t tests, Mann-Whitney U tests and χ2 tests, respectively. Results:The age of 231 IHC cases was 43 (38, 51) years old, with 95.2% (220/231) aged ≥30 years, and males accounted for 64.9% (150/231). HBsAg and HBV DNA levels were 131.9 (20.8, 400.9) IU/mL and 94.0 (0, 448.5) IU/mL, respectively, of which 35.9% (83/231) were HBV DNA negative (<20 IU/mL). The remarkable proportions of G≥2, S≥2, and liver injury (G≥2 and/or S≥2) in liver tissue were 16.5% (38/231), 29% (67/231), and 35.9% (83/231), respectively. The S≥2 proportion was significantly higher in the HBV DNA-negative group than the positive group (42.2% vs. 21.6%, P<0.001), and it mainly occurred in the population cohort over 30 years old (44.9% vs. 31.0%, P=0.04). The liver stiffness measurement (LSM), aspartate transaminase to platelet ratio index (APRI), and platelet (PLT) were significantly higher in the S≥2 group than the S<2 group ( P<0.05). Conclusion:Clinicians can comprehensively evaluate the degree of liver fibrosis in IHC based on clinical factors such as age, PLT, APRI, and LSM, even if the liver histological results are lacking. The China 2022 version guidelines define that nearly half of IHC has histological indications for antiviral therapy, and liver biopsy and prompt treatment can be recommended.
7.Correlation between the ratio of tumor volume to uterine volume and the expression of Ki-67 and p16 pro-tein in tissues with the pathological features and recurrence of endometrial carcinoma
Liping CHEN ; Juyu LUO ; Zhangyan PENG ; Xiulan WU ; Yuhong YANG ; Lianyan SHI ; Xiaoyun LI ; Ling WANG
The Journal of Practical Medicine 2024;40(23):3367-3372
Objective To investigate the correlation between the ratio of lesion volume to uterine volume(T/U),the expression levels of Ki-67 and p16 proteins in lesion tissue,and the recurrence risk of endometrial cancer.Methods A total of 150 patients diagnosed with endometrial carcinoma through pathological examination at Qiandongnan Prefecture People's Hospital were enrolled for follow-up observation.Among them,28 patients experienced recurrence after a 2-year follow-up period,while 122 patients remained recurrence-free.The expression differences of Ki-67 protein and p16 protein in T/U and lesion tissues during surgery were compared between the two groups.Furthermore,these indexes were analyzed based on different pathological features,and the variation in relapse-free survival time was assessed among patients with distinct T/U status as well as Ki-67 and p16 protein expressions.Results The T/U value and the positive expression rate of Ki-67 protein were significantly higher in the relapsed group compared to the non-relapsed group,while the positive expression rate of p16 protein was significantly lower in the relapsed group(P<0.05).Additionally,patients with T/U ≥ 0.18 had a significantly higher proportion of stage Ⅲ patients and patients with low histological differentiation compared to those with T/U<0.18(P<0.05).Furthermore,patients with positive expression of Ki-67 protein exhibited a significantly higher proportion of stage Ⅲ patients,patients with low histological differentiation,and lymph node metastasis compared to those with negative expression of Ki-67 protein(P<0.05).The proportion of stage Ⅲ patients exhibiting positive p16 protein expression,low histological differentiation,and lymph node metastasis was significantly lower compared to those with negative p16 protein expression(P<0.05).Patients with endometrial cancer having a T/U ≥0.18 experienced shorter recurrence-free survival time 2 years post-surgery in comparison to patients with T/U<0.18(x2=6.962,P=0.008).Patients displaying positive Ki-67 expression had a shorter recurrence-free survival time 2 years after surgery than those with negative Ki-67 expression(x2=4.815,P=0.028).The recurrence-free survival time 2 years after surgery for patients expressing p 16 protein positively was longer than that for patients ex-pressing it negatively(x2=4.279,P=0.039).The presence of FIGO stage Ⅲ,lymph node metastasis,depth of myographic invasion ≥1/2,T/U value ≥ 0.18,and positive expression of Ki-67 protein were identified as significant risk factors for postoperative recurrence in endometrial cancer(P<0.05).Conversely,the positive expression of p16 protein was found to be a protective factor against recurrence in endometrial carcinoma following surgery(P<0.05).Conclusion The expression of T/U,Ki-67 protein,and p16 protein in endometrial cancer patients is as-sociated with tumor progression and may augment the risk of postoperative recurrence.
8.Recombinant Schistosoma japonicum cystatin alleviates acute liver injury in mice by inhibiting endoplasmic reticulum stress,inflammation and hepatocyte apoptosis
Lingjun LU ; Xiaodi YANG ; Huaping ZHANG ; Yuan LIANG ; Xiulan SHI ; Xin ZHOU
Journal of Southern Medical University 2024;44(6):1126-1134
Objective To investigate the protective effect of recombinant Schistosoma japonicum cystatin(rSj-Cys)against acute liver injury induced by lipopolysaccharide(LPS)and D-GalN in mice.Methods Adult male C57BL/6J mice with or without LPS/D-GaIN-induced acute liver injury were given intraperitoneal injections of rSj-Cys or PBS 30 min after modeling(n=18),and serum and liver tissues samples were collected from 8 mice in each group 6 h after modeling.The survival of the remaining 10 mice in each group within 24 h was observed.Serum levels of ALT,AST,TNF-α and IL-6 of the mice were measured,and liver pathologies was observed with HE staining.The hepatic expressions of macrophage marker CD68,Bax,Bcl-2 and endoplasmic reticulum stress(ERS)-related proteins were detected using immunohistochemistry or immunoblotting,and TUNEL staining was used to detect hepatocyte apoptosis.Results The survival rates of PBS-and rSj-Cys-treated mouse models of acute liver injury were 30%and 80%at 12 h and were 10%and 60%at 24 h after modeling,respectively;no death occurred in the two control groups within 24 h.The mouse models showed significantly increased serum levels of AST,ALT,IL-6 and TNF-α and serious liver pathologies with increased hepatic expressions of CD68 and Bax,lowered expression of Bcl-2,increased hepatocyte apoptosis,and up-regulated expressions of ERS-related signaling pathway proteins GRP78,CHOP and NF-κB p-p65.Treatment of the mouse models significantly lowered the levels of AST,ALT,IL-6 and TNF-α,alleviated liver pathologies,reduced hepatic expressions of CD68,Bax,GRP78,CHOP and NF-κB p-p65,and enhanced the expression of Bcl-2.In the normal control mice,rSj-Cys injection did not produce any significant changes in these parameters compared with PBS.Conclusion rSj-Cys alleviates LPS/D-GalN-induced acute liver injury in mice by suppressing ERS,attenuating inflammation and inhibiting hepatocyte apoptosis.
9.Correlation between serum lncRNA FGD5-AS1,miR-103a-3p and puerperal infection in patients with gestational diabetes mellitus in late pregnancy
Suying WANG ; Yingxin HAN ; Xiulan CHENG ; Yanqing LI ; Jing ZHAO ; Chunhong YANG ; Chunyan ZHANG
International Journal of Laboratory Medicine 2024;45(14):1720-1724
Objective To investigate the correlation between serum long non-coding RNA FGD5-AS1(ln-cRNA FGD5-AS1),microRNA-103a-3p(miR-103a-3p)and puerperal infection(PI)in patients with gesta-tional diabetes mellitus(GDM)in late pregnancy.Methods A total of 168 late pregnancy GDM patients who were hospitalized and delivered in the hospital from January 2022 to June 2023 were retrospectively selected as the experimental group,and the patients were separated into an infected group(96 cases)and an uninfected group(72 cases)based on whether they had PI.At the same time,120 late pregnant women who underwent prenatal examination in the hospital and had normal gestational blood glucose were selected as the control group.Real-time fluorescence quantitative PCR(qRT-PCR)was applied to detect the expression levels of ln-cRNA FGD5-AS1 and miR-103a-3p.Multivariate Logistic regression was applied to analyze the influencing factors of PI in late pregnancy GDM patients.StarBase website was applied to analyze the relationship between lncRNA FGD5-AS1 and miR-103a-3p.Pearson was applied to analyze the correlation between lncRNA FGD5-AS1 and miR-103a-3p.Receiver operating characteristic(ROC)curve was applied to evaluate the value of ln-cRNA FGD5-AS1 and miR-103a-3p in predicting the occurrence of PI.Results There was a statistically sig-nificant difference in the expression levels of serum lncRNA FGD5-AS1 and miR-103a-3p between the experi-mental group and the control group(P<0.05),the expression level of serum lncRNA FGD5-AS1 in the infec-ted group was obviously higher than that in the uninfected group(P<0.05),but the expression level of ser-um miR-103a-3p in the infected group was obviously lower than that in the uninfected group(P<0.05).The expression level of lncRNA FGD5-AS1 was an independent risk factor for PI in late-pregnancy GDM patients(P<0.05),and the expression level of miR-103a-3p was an independent protective factor for PI in late-preg-nancy GDM patients(P<0.05).There was a negative correlation between lncRNA FGD5-AS1 and miR-103a-3p expression level(r=-0.409,P<0.001).The efficacy of the combined detection of lncRNA FGD5-AS1 and miR-103a-3p for predicting PI in late pregnancy GDM patients was superior to that of serum lncRNA FGD5-AS1 and miR-103a-3p alone(P<0.05).Conclusion LncRNA FGD5-AS1 is an independent risk factor for PI in late pregnancy GDM patients,while miR-103a-3p is an independent protective factor for PI in late pregnancy GDM patients.The combined detection has higher value for predicting PI in late pregnancy GDM patients.
10.Exploration and practice of humanistic care in children's fever clinics from the perspective of narrative medicine
Yutao CAI ; Xue CHEN ; Xiulan WEN ; Liyan YE ; Handan HONG ; Canying LI ; Xian CAI ; Qiumei YANG ; Yansong SU
Chinese Medical Ethics 2024;37(5):586-591
To explore the application of humanistic care in children's fever clinics,the children's fever clinics of Guangzhou Women and Children's Medical Center conducted relevant research using narrative medicine as a guiding concept.This paper elaborated on the medical humanistic dilemmas and narrative care needs in children's fever clinics,and focused on exploring the practice paths of narrative medicine in pediatric with Chinese characteristics from five dimensions,including the cultivation of nursing staff's abilities of professional narrative and humanistic care,the establishment of health lectures featuring narrative patient education,the alleviation of medical anxiety for children and their families,related support of narrative nursing,and caring services.The aim was to improve the narrative care ability of nurses in children's fever clinics,develop methods for pediatric patients that can eliminate the fear of seeking medical treatment,and protect children's physical and mental health.A carrier of care,support,and emotional expression can be provided for parents.The foundation for nursing staff to achieve professional growth through narrative reflection can be laid.Thus,it can assist in establishing a life narrative community relationship between doctors and patients,and jointly explore the meaning of life.

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