1.Mechanistic Study on Tougu Xiaotong Capsules in Regulating PANoptosis to Delay Degeneration of Chondrocytes in Knee Osteoarthritis
Jinxia YE ; Yixin LIN ; Xiaoqing LEI ; Yanfeng HUANG ; Changlong FU ; Desen LI ; Wenyi WANG ; Lan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):149-161
ObjectiveTo investigate the effect of Tougu Xiaotong capsules (TGXTC) on the regulation of chondrocyte PANoptosis, delay of chondrocyte degeneration, and improvement of the symptoms in knee osteoarthritis (KOA). MethodsIn vivo experiments: 50 male C57BL/6 mice were randomly assigned into five groups (n=10 per group): sham operation group, model group, low-dose TGXTC group (7.2 g·kg-1), high-dose TGXTC group (14.4 g·kg-1), and diclofenac sodium group (0.05 g·kg-1). Except for the sham group, KOA models were established in all other groups using the modified Hulth method. Following successful model induction, the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg-1 for 6 weeks, while the diclofenac sodium group received 0.05 g·kg-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score; micro-computed tomography (micro-CT) was used to scan the knee, hematoxylin-eosin (HE) staining and safranin O-fast green staining were used to observe the morphology of cartilage, transmission electron microscopy (TEM) was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence (IF) co-localization assays was performed to detect the co-localization of cleaved Caspase-3, receptor-interacting protein 3 (RlPK3), and the N-terminal domain of gasdermin D (GSDMD-N) in cartilage tissue, while western blot was employed to detect the expression levels of cleaved Caspase-3, RIPK3, and GSDMD-N. In vitro experiments: The knee cartilages of 4-week-old SD rats were isolated, and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups: the control group, the model group (pretreated with 10 mg·L-1 lipopolysaccharide (LPS) for 24 h followed by treatment with 1 μmol·L-1 nigericin for 4 h), and the TGXTC treatment group (pretreated with 10 mg·L-1 LPS for 24 h, followed by exposure to 1 μmol·L-1 nigericin for 4 h and subsequently treated with 100 mg·L-1 TGXTC for an additional 24 h). The levels of reactive oxygen species (ROS), apoptosis, necroptosis, and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection, AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels (IL-1β and IL-18) were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis, including cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3. ResultsCompared with the sham group, the Lequesne MG scores were significantly up-regulated(P<0.01) in the model group, and the pathological changes of cartilage were significantly, with joint spaces narrower, osteophyte formation increased, secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis, necroptosis, and pyroptosis, along with markedly elevated expression of cleaved Caspase-3, RlPK3, and GSDMD-N in cartilage tissue (P<0.01). In addition, The mean fluorescence intensities of ROS, orange-red fluorescence in AO/EB staining, green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group (P<0.01) . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased (P<0.01). The expression of PANoptosis related proteins (cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3) were also significantly upregulated(P<0.05). Compared to the model group, the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score, an increase in joint space, a decrease in osteophyte formation, diminished cartilage damage, reduced release of ROS, and alleviation of apoptotic, necroptotic, and pyroptotic processes in chondrocytes. Additionally, mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased (P<0.05). Furthermore, the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions (P<0.05). Similar results were observed in chondrocytes: cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3 exhibited significant decreases as well (P<0.05). ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.
2.Mechanistic Study on Tougu Xiaotong Capsules in Regulating PANoptosis to Delay Degeneration of Chondrocytes in Knee Osteoarthritis
Jinxia YE ; Yixin LIN ; Xiaoqing LEI ; Yanfeng HUANG ; Changlong FU ; Desen LI ; Wenyi WANG ; Lan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):149-161
ObjectiveTo investigate the effect of Tougu Xiaotong capsules (TGXTC) on the regulation of chondrocyte PANoptosis, delay of chondrocyte degeneration, and improvement of the symptoms in knee osteoarthritis (KOA). MethodsIn vivo experiments: 50 male C57BL/6 mice were randomly assigned into five groups (n=10 per group): sham operation group, model group, low-dose TGXTC group (7.2 g·kg-1), high-dose TGXTC group (14.4 g·kg-1), and diclofenac sodium group (0.05 g·kg-1). Except for the sham group, KOA models were established in all other groups using the modified Hulth method. Following successful model induction, the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg-1 for 6 weeks, while the diclofenac sodium group received 0.05 g·kg-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score; micro-computed tomography (micro-CT) was used to scan the knee, hematoxylin-eosin (HE) staining and safranin O-fast green staining were used to observe the morphology of cartilage, transmission electron microscopy (TEM) was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence (IF) co-localization assays was performed to detect the co-localization of cleaved Caspase-3, receptor-interacting protein 3 (RlPK3), and the N-terminal domain of gasdermin D (GSDMD-N) in cartilage tissue, while western blot was employed to detect the expression levels of cleaved Caspase-3, RIPK3, and GSDMD-N. In vitro experiments: The knee cartilages of 4-week-old SD rats were isolated, and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups: the control group, the model group (pretreated with 10 mg·L-1 lipopolysaccharide (LPS) for 24 h followed by treatment with 1 μmol·L-1 nigericin for 4 h), and the TGXTC treatment group (pretreated with 10 mg·L-1 LPS for 24 h, followed by exposure to 1 μmol·L-1 nigericin for 4 h and subsequently treated with 100 mg·L-1 TGXTC for an additional 24 h). The levels of reactive oxygen species (ROS), apoptosis, necroptosis, and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection, AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels (IL-1β and IL-18) were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis, including cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3. ResultsCompared with the sham group, the Lequesne MG scores were significantly up-regulated(P<0.01) in the model group, and the pathological changes of cartilage were significantly, with joint spaces narrower, osteophyte formation increased, secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis, necroptosis, and pyroptosis, along with markedly elevated expression of cleaved Caspase-3, RlPK3, and GSDMD-N in cartilage tissue (P<0.01). In addition, The mean fluorescence intensities of ROS, orange-red fluorescence in AO/EB staining, green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group (P<0.01) . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased (P<0.01). The expression of PANoptosis related proteins (cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3) were also significantly upregulated(P<0.05). Compared to the model group, the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score, an increase in joint space, a decrease in osteophyte formation, diminished cartilage damage, reduced release of ROS, and alleviation of apoptotic, necroptotic, and pyroptotic processes in chondrocytes. Additionally, mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased (P<0.05). Furthermore, the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions (P<0.05). Similar results were observed in chondrocytes: cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3 exhibited significant decreases as well (P<0.05). ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.
3.Clinical value and safety of endoscopic-assisted skin-sparing mastectomy combined with immediate implant-based breast reconstruction as day surgery for breast cancer
Youxing FU ; Xiaoqing LONG ; Zhongjian ZHU ; Mingjun HUANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(01):37-43
Objective To investigate the feasibility, safety, and clinical value of endoscopic-assisted skin-sparing mastectomy combined with immediate implant-based breast reconstruction performed as day surgery for breast cancer, aiming to provide a reference for major hospitals seeking to implement a day surgery model for breast cancer treatment. Methods We retrospectively analyzed the patients who underwent endoscopic-assisted skin-sparing mastectomy combined with immediate implant-based breast reconstruction for breast cancer at West China Hospital of Sichuan University from June 2021 to December 2022, and they were divided into a day surgery group and a conventional inpatient group based on their admission model. The operative indicators, Breast-Q scores, preoperative waiting time, length of hospital stay, hospitalization costs and complications of the two groups were analyzed. Results Except for intraoperative bleeding (P=0.007), the difference between the two groups in comparison of the rest of the operative indicators was not statistically significant (all P>0.05); there was no significant difference between the two groups in preoperative and postoperative Breast-Q scores (all P>0.05); the preoperative waiting time and length of stay in hospital of the day surgery group were 4.0 (3.0, 11.0) days and 1.0 (1.0, 1.0) days, respectively, which were significantly shorter than that of the conventional inpatient group; the postoperative pain score in the day surgery group [1.0 (1.0, 1.0) points] was lower than that in the conventional inpatient group [3.0 (3.0, 3.0) points], with a statistically significant difference between the two groups (P<0.001). Additionally, the total hospitalization costs for the day surgery group and conventional inpatient group were 50 656.5 (48 145.3, 62 597.3) RMB and 53 689.3 (50 469.1, 64 826.5) RMB, respectively.The total hospitalization cost in the day surgery group was significantly lower than that in the conventional inpatient group, with a statistically significant difference between the two groups (P=0.001). There was no statistically significant difference in complications between the two groups (all P>0.05). Conclusion Endoscopic-assisted skin-sparing mastectomy combined with immediate implant-based breast reconstruction in day surgery is feasible and safe. Without increasing postoperative complications, it effectively reduces hospitalization costs and shortens medical care time, demonstrating significant clinical value.
4.Whole genomic analysis of 8 strains of H9N2 subtype avian influenza virus isolates from live poultry markets in Yunnan, 2023
LIU Zhaosheng ; FU Xiaoqing ; LUO Chunrui
China Tropical Medicine 2025;25(3):350-
Objective To conduct an in-depth study of the molecular biological characteristics and evolutionary trends of H9N2 avian influenza virus (AIV) in live poultry markets in Yunnan Province in 2023, and to provide scientific evidence for the development of control strategies for H9N2 avian influenza in the region. Methods Environmental samples were collected from live poultry markets in Yunnan Province in 2023 for H9N2 subtype nucleic acid detection. Positive samples were subjected to virus isolation using chicken embryos, and the genome of the 8 isolated strains was amplified, sequenced, and analyzed for genetic characteristics. Results The eight avian influenza virus (AIV) isolates had the hemagglutinin (HA) cleavage site sequence PSRSSRGLF, which is a non-continuous basic amino acid sequence, consistent with the genetic characteristics of typical low-pathogenicity avian influenza viruses. Mutations Q234L and H191N were observed in the left arm of the HA protein, which enhanced the affinity for α-2,6 sialic acid receptors, suggesting that these viruses may have the potential to infect humans. The neuraminidase (NA) protein exhibited a deletion of three amino acids (TEI) at positions 62–64 in the stalk region, displaying characteristics of high pathogenicity at the molecular level. The increase or absence in potential glycosylation sites were observed in both HA and NA genes. The non-structural protein 1 (NS1) showed no D92E mutation, and had a C-terminal truncation of 13 amino acids, indicating that this virus is of low pathogenicity and poses a lower risk of human transmission. Mutations T37A, R95K, S224N, and K242N in the M1 protein of some isolates increased the risk of infection, while one isolate carried the V27A or S31N mutation in the M2 protein, conferring resistance to M2 ion channel inhibitors. Mutations M317I and S678N were identified in the PB1 protein, which may enhance pathogenicity in mice and increase the potential for mammalian infection. The PB2 protein carried the I292V mutation, which exhibited a stronger infectivity to mammals. Phylogenetic analysis revealed that the HA, NA, and PB2 gene segments belonged to the Y280 lineage, NP and PB1 gene segments were classified under the F98 lineage, the M gene segment of the NH013 isolate belonged to the F98 lineage, while M genes, as well as the NS and PA genes of other isolates belonged to the G1 lineage. Conclusions These eight AIV isolates exhibited characteristics of low pathogenicity, but simultaneously carry the potential risk of infecting humans. Despite the HA cleavage site and NS1 protein mutations indicating low pathogenicity, the Q234L and H191N mutations in the HA protein enhanced its affinity for human receptors, suggesting the potential for human infection. The TEI deletion in the NA protein, mutations in the M1 protein, and resistance mutations in the M2 protein further increase the risk of human infection. Mutations in the PB1 and PB2 proteins increase the potential for these eight AIV strains to infect humans or mammals.
5.Efficacy of virtual reality technology in patients with schizophrenia treatment
Qing YUAN ; Dan LIU ; Xiaoqing FU ; Tianhao BAO
Sichuan Mental Health 2024;37(3):277-281
Schizophrenia is a severe mental illness primarily managed with medication.In recent years,with the rapid development of virtual reality(VR)technology,its application effect in mental illness has been widely concerned.This review aims to explore the application of VR technology in schizophrenia treatment and to provide references for clinical practice.By reviewing randomized controlled trials from both domestic and international sources,the therapeutic efficacy of VR technology in treating schizophrenia was evaluated.The findings consistently demonstrated that VR technology has a positive effect on hallucinations,cognitive function,stress management and emotional control,and social function recovery in schizophrenia.
6.Current status and reflection on molecular diagnosis of birth defects
Ming QI ; Qihua FU ; Xiaoqing ZHANG
Chinese Journal of Preventive Medicine 2024;58(2):166-170
Birth defects are a major problem threatening the health of children in China. Genetic factors play a major role in birth defect etiology. Molecular diagnosis is the key means for screening, diagnosing, and preventing birth defects caused by genetic factors. How to carry out large-scale and cost-effective molecular diagnosis in clinical practice is a major challenge in the prevention and treatment of birth defects in China. This article reviews the current status of birth defects in China, the application of molecular diagnostic technology in birth defect prevention and control, and the challenges in promoting its use, to provide references for clinical practice in birth defect molecular diagnosis.
7.Study of gemcitabine in end-stage liver disease with sepsis
Rui WU ; Yunlei XIAO ; Xiaoqing FU
China Modern Doctor 2024;62(20):104-107,112
Objective To observe the effect and prognosis of recombinant human granulocyte colony-stimulating factor(rhG-CSF)in end-stage liver disease(ESLD)with sepsis.Methods Ninety patients with ESLD complicated with sepsis from January 2022 to December 2023 were selected and randomly divided into rhG-CSF group,thymosin group and control group,with 30 cases each.Compare the liver function,cytokine levels,treatment efficacy,complications,and ESLD model(MELD)scoring system before and after treatment among three groups.Results The total effective rate of rhG-CSF group was 96.67%,which was significantly higher than the other two groups,and the difference was statistically significant(P<0.05).After treatment,the liver function,cytokine levels,and immune function recovery of the rhG-CSF group and thymosin group were better than that of control group,with statistical significance(P<0.05).After 12 weeks of treatment and follow-up,the least complications and the lowest mortality rate in the rhG-CSF group.Conclusion The clinical efficacy of rhG-CSF as an adjuvant therapy for ESLD complicated with sepsis is excellent,which can enhance anti-infection ability and improve prognosis.
8.Analysis of quality of life and influencing factors in patients with multidrug-resistant tuberculosis
Yajun SHI ; Xiaoqin XIONG ; Xiaoqing FU ; Feng WANG ; Hui ZHOU
China Modern Doctor 2024;62(29):10-14
Objective To explore the health-related quality of life(HRQOL)among patients with multidrug-resistant and rifampin-resistant pulmonary tuberculosis(MDR/RR-PTB)and analyze its influencing factors.Methods Data was collected from 50 MDR/RR-PTB patients registered for treatment management in the"Tuberculosis Management Information System"in Bazhong City from 2021 to 2024 as resistance group.Fifty pulmonary tuberculosis patients sensitive to anti-tuberculosis drug treatment registered in the system during the same period were selected as control group.A cross-sectional survey method was employed using the quality of life instruments for chronic disease pulmonary tuberculosis(QLICD-PT)to measure and compare the HRQOL between two groups.Multiple linear regression analysis was used to analyze factors affecting the HRQOL of MDR/RR-PTB patients.Results Except for physiological function,there were statistically significant difference between MDR/RR-PTB group and control group in terms of total quality of life score,psychological function,social function,and specific modules(P<0.05).Univariate analysis showed that there were statistically significant differences in the total quality of life scores among patients with different genders,medical insurance statuses,monthly family income situations,presence or absence of comorbidities,lymphocyte count,albumin,C-reactive protein,and body mass index(P<0.05).Multiple linear regression analysis revealed that(P<0.05)there was significant difference for the partial regression coefficient tests of patient's medical insurance and monthly family income situations(P<0.05).Conclusion The HRQOL of MDR/RR-PTB patients is lower than that of pulmonary tuberculosis patients sensitive to anti-tuberculosis drugs.The patient's health insurance and monthly family income are potential factors affecting the HRQOL of MDR/RR-PTB patients.
9.Factors influencing advance care planning engagement behavior in patients with advanced cancer:a systematic review
Yiling FU ; Qian WU ; Xiaoqing LUO ; Aihong WU ; Xuelan XIA ; Min ZHENG
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(4):482-493
Objective·To systematically integrate relevant influencing factors of advanced cancer patients'engagement behavior in advance care planning(ACP).Methods·The systematic search of Chinese and English literature on factors influencing ACP engagement behavior in advanced cancer patients from inception to December 2022 in China National Knowledge Infrastructure(CNKI),Wanfang,China Biomedical Literature Database(Sinomed),PubMed,Cochrane Library,Embase,CINAHL,and PsycINFO was conducted.After the literature quality evaluation,content extraction and summary were conducted by two researchers,and the data of quantitative research and qualitative research were extracted and integrated respectively.The final influencing factors of ACP engagement behavior of advanced cancer patients were obtained.With the help of the theoretical domain,they were mapped to the capability,opportunity,motivation-behavior(COM-B)model step by step.Results·A total of 21 studies were included and 27 factors were summarized,including 9 theoretical domains.Mapping to the COM-B model included 9 capability factors(knowledge of ACP,education level,accurate knowledge of prognosis,knowledge of the time of disease diagnosis,prior experience,subjective life expectancy,age,cancer site,and disease symptom burden),13 opportunity factors(gender,marital status,race/ethnicity,religious belief,dependent children,family economic condition,place of living,housing type,family support,social support,doctor-patient relationship,acculturation,and whether or not to establish a hospice service center)and 5 motivational factors(ACP attitude,ACP belief,ACP motivation,anxiety and depression,and death attitude).Among them,doctor-patient relationship,religious belief,ACP attitude,educational level,marital status,family support,knowledge of ACP,accurate knowledge of prognosis,age,place of living,attitude toward death,prior experience,and race/ethnicity were more influential factors on ACP engagement behavior.Conclusion·Based on the COM-B model,the influencing factors of ACP engagement behavior in advanced cancer patients can be comprehensively summarized.Future studies can use the above factors as an entry point to design continuous,multifaceted,and comprehensive interventions based on the COM-B model to promote the practice of ACP engagement behavior in advanced cancer patients.
10.Effects of Natural Moxibustion at "Feishu (BL 13)" on Th17/Treg Immune Balance and Inflammatory Response in Healthy Rats
Jia LIN ; Xiaomei SHAO ; Ruiyang FU ; Jinxia LI ; Xiaoqing GUO
Journal of Traditional Chinese Medicine 2024;65(16):1703-1708
ObjectiveTo observe the effect of natural moxibustion at "Feishu (BL 13)" on immune balance of T helper cell 17 (Th17) / regulatory T cell (Treg) in healthy rats. MethodsForty-eight rats were randomly divided into 10 rats in the sham moxibustion group and 38 rats in natural moxibustion group. The rats in the sham moxibustion group applied blank acupoint stickers to bilateral "Feishu (BL 13)", and the rats in natural moxibustion group applied acupoint stickers filled with Compound Banmao Ointment (复方斑蝥膏) to bilateral "Feishu (BL 13)" for a period of 8 h. Thirty rats in natural moxibustion group were successfully blistered after 8 h, and then were randomly divided into 1-day, 3-day and 7-day groups with 10 rats in each group. The general condition of rats was recorded during the experiment; different time groups of natural moxibustion were sampled at the corresponding time, and HE staining was used to observe the pathological changes of the skin in the area of application; flow cytometry was used to detect the subpopulations of Th17 and Treg in peripheral blood, and the value of Th17/Treg was calculated; and ELISA was used to detect the serum interleukin 17A (IL-17A) and interleukin 6 (IL-6), interleukin 10 (IL-10) levels. ResultsCompared with sham moxibustion group, blisters can be seen in the application area of rats in 1-day natural moxibustion group, and the rats often scratched the skin of the moxibustion area, which showed loose stratum corneum, thickening of the stratum spinosum and stratum granulosum, absence of cells in the basal layer, inflammatory cell infiltration, and elevation of Treg in peripheral blood, and serum IL-17A, IL-6; in 3-day natural moxibustion group, the moxibustion area of the rats was scabbed and partially detached, with the most obvious dermatopathological changes, and elevated peripheral blood Th17 and Treg, and serum IL-17A, IL-6, IL-10; in 7-day natural moxibustion group, skin damage and pathological changes in the area of moxibustion were basically restored; Th17/Treg values were reduced in 1-, 3- and 7-day moxibustion groups after blistering (P<0.05 or P<0.01). Compared with the 1-day moxibustion group, peripheral blood Th17, Treg, and serum IL-17A elevated in the 3-day moxibustion group; peripheral blood Treg, serum IL-17A, and IL-6 decreased, and Th17/Treg values elevated in the 7-day moxibustion group (P<0.05 or P<0.01). Compared with the 3-day moxibustion group, the 7-day moxibustion group had lower peripheral blood Th17 and Treg, serum IL-17A, IL-6, and IL-10, and higher Th17/Treg values (P<0.05 or P<0.01). ConclusionNatural moxibustion at "Feishu (BL 13)" can shift the Th17/Treg balance towards Treg of healthy rats, which will gradually lead to immune homeostasis, and regulate the relevant inflammatory factors in the serum to prevent inflammation from occurring and developing.

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