1.Status of social support and influencing factors of support utilization among general practitioners in Minhang District of Shanghai
Shuai LIU ; Na XU ; Liqun GAO ; Qingzhen LONG ; Yonghong MU ; Weiying GU
Chinese Journal of General Practitioners 2025;24(3):288-295
Objective:To analyze the status of social support and the influencing factors of support utilization among general practitioners.Methods:An investigation on the status of social support and the influencing factors of support utilization was conducted among general practitioners (GPs) from 6 communities in Shanghai Minhang District selected by simple sampling method in February 2023. The investigation included a questionnaire survey containing basic information and intervention measures (emotional support, information support, tool support); and assessments of the Social Support Rate Scale (SSRS), Perceived Social Support Scale, General Self-Efficacy Scale, Quality of Work Life Scale, Trait Coping Style Scale, and Psychological Resilience Scale. The association of social support status with various characteristics of GPs and their perceived social support and trait coping style was analyzed, and the influencing factors of social support utilization were determined.Results:A total of 184 questionnaires were distributed and 184 valid questionnaires were collected, with an effective recovery rate of 100.00%. Among the 184 participants, 55 (29.89%) had a general level of social support, and 129 (70.11%) were satisfied with their social support. The total score of SSRS was (37.49±8.41), with the objective support dimension scoring (9.67±3.42), subjective support scoring (20.04±4.52), and support utilization scoring (7.77±2.09). Univariate analysis showed significant differences in social support levels among respondents with different marital status, number of children, emotional support, informational support, instrumental support, positive coping, negative coping, psychological resilience, compassion fatigue, general self-efficacy, and perceived social support status ( P<0.05). Multiple linear regression analysis indicated that marital status, family support, professional skills, positive coping, and negative coping were independent influencing factors of social support. Among these factors, being married, having family care support, professional titles and skills and positive coping had a positive impact on social support levels, while negative coping had a negative impact ( P<0.05). Further stepwise multiple linear regression analysis showed that friend support, positive coping and family care were positive influencing factors for support utilization, while negative coping was a negative influencing factor ( P<0.05). Conclusion:The levels of social support among GPs in Minghang district of Shanghai are relatively high. Marital status, family support, professional title and skills and positive coping are positive factors for social support; and friend support, positive coping and family support are positive factors for support utilization.
2.Study on the mechanistic role of the Fuzheng Huayu formula against cholestatic liver fibrosis in mice
Zheng ZHANG ; Yue LIANG ; Enqi TANG ; Xiaoxi ZHOU ; Yonghong HU ; Gaofeng CHEN ; Wei LIU ; Yongping MU ; Ping LIU ; Jiamei CHEN
Chinese Journal of Hepatology 2025;33(9):889-897
Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2-/ -) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2-/ -mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 -/ - mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 -/ - mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 -/ - mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.
3.Status of social support and influencing factors of support utilization among general practitioners in Minhang District of Shanghai
Shuai LIU ; Na XU ; Liqun GAO ; Qingzhen LONG ; Yonghong MU ; Weiying GU
Chinese Journal of General Practitioners 2025;24(3):288-295
Objective:To analyze the status of social support and the influencing factors of support utilization among general practitioners.Methods:An investigation on the status of social support and the influencing factors of support utilization was conducted among general practitioners (GPs) from 6 communities in Shanghai Minhang District selected by simple sampling method in February 2023. The investigation included a questionnaire survey containing basic information and intervention measures (emotional support, information support, tool support); and assessments of the Social Support Rate Scale (SSRS), Perceived Social Support Scale, General Self-Efficacy Scale, Quality of Work Life Scale, Trait Coping Style Scale, and Psychological Resilience Scale. The association of social support status with various characteristics of GPs and their perceived social support and trait coping style was analyzed, and the influencing factors of social support utilization were determined.Results:A total of 184 questionnaires were distributed and 184 valid questionnaires were collected, with an effective recovery rate of 100.00%. Among the 184 participants, 55 (29.89%) had a general level of social support, and 129 (70.11%) were satisfied with their social support. The total score of SSRS was (37.49±8.41), with the objective support dimension scoring (9.67±3.42), subjective support scoring (20.04±4.52), and support utilization scoring (7.77±2.09). Univariate analysis showed significant differences in social support levels among respondents with different marital status, number of children, emotional support, informational support, instrumental support, positive coping, negative coping, psychological resilience, compassion fatigue, general self-efficacy, and perceived social support status ( P<0.05). Multiple linear regression analysis indicated that marital status, family support, professional skills, positive coping, and negative coping were independent influencing factors of social support. Among these factors, being married, having family care support, professional titles and skills and positive coping had a positive impact on social support levels, while negative coping had a negative impact ( P<0.05). Further stepwise multiple linear regression analysis showed that friend support, positive coping and family care were positive influencing factors for support utilization, while negative coping was a negative influencing factor ( P<0.05). Conclusion:The levels of social support among GPs in Minghang district of Shanghai are relatively high. Marital status, family support, professional title and skills and positive coping are positive factors for social support; and friend support, positive coping and family support are positive factors for support utilization.
4.Study on the mechanistic role of the Fuzheng Huayu formula against cholestatic liver fibrosis in mice
Zheng ZHANG ; Yue LIANG ; Enqi TANG ; Xiaoxi ZHOU ; Yonghong HU ; Gaofeng CHEN ; Wei LIU ; Yongping MU ; Ping LIU ; Jiamei CHEN
Chinese Journal of Hepatology 2025;33(9):889-897
Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2-/ -) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2-/ -mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 -/ - mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 -/ - mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 -/ - mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.
5.A multicenter study on effect of delayed chemotherapy on prognosis of Burkitt lymphoma in children
Li SONG ; Ling JIN ; Yonghong ZHANG ; Xiaomei YANG ; Yanlong DUAN ; Mincui ZHENG ; Xiaowen ZHAI ; Ying LIU ; Wei LIU ; Ansheng LIU ; Xiaojun YUAN ; Yunpeng DAI ; Leping ZHANG ; Jian WANG ; Lirong SUN ; Rong LIU ; Baoxi ZHANG ; Lian JIANG ; Huixia WEI ; Kailan CHEN ; Runming JIN ; Xige WANG ; Haixia ZHOU ; Hongmei WANG ; Shushuan ZHUANG ; Chunju ZHOU ; Zifen GAO ; Xiao MU ; Kaihui ZHANG ; Fu LI
Chinese Journal of Pediatrics 2024;62(10):941-948
Objective:To analyze the factors affecting delayed chemotherapy in children with Burkitt lymphoma (BL) and their influence on prognosis.Methods:Retrospective cohort study. Clinical data of 591 children aged ≤18 years with BL from May 2017 to December 2022 in China Net Childhood Lymphoma (CNCL) was collected. The patients were treated according to the protocol CNCL-BL-2017. According to the clinical characteristics, therapeutic regimen was divided into group A, group B and group C .Based on whether the total chemotherapy time was delayed, patients were divided into two groups: the delayed chemotherapy group and the non-delayed chemotherapy group. Based on the total delayed time of chemotherapy, patients in group C were divided into non-delayed chemotherapy group, 1-7 days delayed group and more than 7 days delayed group. Relationships between delayed chemotherapy and gender, age, tumor lysis syndrome before chemotherapy, bone marrow involvement, disease group (B/C group), serum lactate dehydrogenase (LDH) > 4 times than normal, grade Ⅲ-Ⅳ myelosuppression after chemotherapy, minimal residual disease in the interim assessment, and severe infection (including severe pneumonia, sepsis, meningitis, chickenpox, etc.) were analyzed. Logistic analysis was used to identify the relevant factors. Kaplan-Meier method was used to analyze the patients' survival information. Log-Rank was used for comparison between groups.Results:Among 591 patients, 504 were males and 87 were females, the follow-up time was 34.8 (18.6,50.1) months. The 3-year overall survival (OS) rate was (92.5±1.1)%,and the 3-year event-free survival (EFS) rate was (90.5±1.2)%. Seventy-three (12.4%) patients were in delayed chemotherapy group and 518 (87.6%) patients were in non-delayed chemotherapy group. The reasons for chemotherapy delay included 72 cases (98.6%) of severe infection, 65 cases (89.0%) of bone marrow suppression, 35 cases (47.9%) of organ dysfunction, 22 cases (30.1%) of tumor lysis syndrome,etc. There were 7 cases of chemotherapy delay in group B, which were seen in COPADM (vincristine+cyclophosphamide+prednisone+daunorubicin+methotrexate+intrathecal injection,4 cases) and CYM (methotrexate+cytarabine+intrathecal injection,3 cases) stages. There were 66 cases of chemotherapy delay in group C, which were common in COPADM (28 cases) and CYVE 1 (low dose cytarabine+high dose cytarabine+etoposide+methotrexate, 12 cases) stages. Multinomial Logistic regression analysis showed that the age over 10 years old ( OR=0.54,95% CI 0.30-0.93), tumor lysis syndrome before chemotherapy ( OR=0.48,95% CI 0.27-0.84) and grade Ⅲ-Ⅳ myelosuppression after chemotherapy ( OR=0.55,95% CI 0.33-0.91)were independent risk factors for chemotherapy delay.The 3-year OS rate and the 3-year EFS rate of children with Burkitt lymphoma in the delayed chemotherapy group were lower than those in the non-delayed chemotherapy group ((79.4±4.9)% vs. (94.2±1.1)%, (80.2±4.8)% vs. (92.0±1.2)%,both P<0.05). The 3-year OS rate of the group C with chemotherapy delay >7 days (42 cases) was lower than that of the group with chemotherapy delay of 1-7 days (22 cases) and the non-delay group (399 cases) ((76.7±6.9)% vs. (81.8±8.2)% vs. (92.7±1.3)%, P=0.002).The 3-year OS rate of the chemotherapy delay group (9 cases) in the COP (vincristine+cyclophosphamide+prednisone) phase was lower than that of the non-chemotherapy delay group (454 cases) ((66.7±15.7)% vs. (91.3±1.4)%, P=0.005). Similarly, the 3-year OS rate of the chemotherapy delay group (11 cases) in the COPADM1 phase was lower than that of the non-chemotherapy delay group (452 cases) ((63.6±14.5)% vs. (91.5±1.3)%, P=0.001). Conclusions:The delayed chemotherapy was related to the age over 10 years old, tumor lysis syndrome before chemotherapy and grade Ⅲ-Ⅳ myelosuppression after chemotherapy in pediatric BL. There is a significant relationship between delayed chemotherapy and prognosis of BL in children.
6.Perceived social support of medical staffs working in community health service institutions in Shanghai Minhang District
Qi SHEN ; Shuai LIU ; Bin CHEN ; Yonghong MU ; Tongtong WEI
Chinese Journal of General Practitioners 2024;23(7):694-701
Objective:To survey the perceived social support status and influencing factors of medical staffs working in community health service institutions in Shanghai Minhang District .Methods:The study was a cross-sectional study. From September 2022 to January 2023, a survey on the status of perceived social support was conducted among 181 medical staffs working in community health service institutions in Shanghai Minhang District. The self-designed general information questionnaire, perceived social support scale, general self-efficacy scale, quality of professional life scale, and trait coping style scale were used in the survey. The effects of demographic characteristics, general self-efficacy, empathy fatigue and trait coping style on perceived social support status of community medical workers were analyzed.Results:A total of 181 questionnairs were distributed and 178 valid ones returned with a valid recovery rate of 98.3%. In three dimensions of perceived social support, family support scored (17.11±8.87), friend support scored (17.08±8.41), other support scored (19.89±4.09), and the total score of understanding social support was (60.99±11.71). There were 2 cases of low support status (1.12%), 81 cases of intermediate support status (45.51%), and 95 cases of high support status (53.37%). Univariate analysis showed that there were significant differences in perceived social support among participants with different ages, marriage status, majors, professional titles, job satisfaction, positive or negative coping styles, general self-efficacy, and empathy fatigue ( P<0.05). Multivariate logistic regression analysis showed that people aged 40 and above had a higher degree of perceived social support than those under 40 ( OR=2.792, 95% CI: 1.051-7.418, P<0.05); compared with public health professionals, clinical medicine, nursing and other medical technique professionals had a higher degree of perceived social support ( OR=4.024, 95% CI: 1.479-10.953; OR=2.941, 95% CI: 1.031-8.393; OR=3.996, 95% CI: 0.543-29.409, all P<0.05); compared with those who were less satisfied with their self-assessment work, participants who were more satisfied or generally satisfied with their self-assessment work had lower perceived social support ( OR=0.200, 95% CI: 0.051-0.781; OR=0.320, 95% CI: 0.146-0.701, all P<0.05); compared with those with mild and moderate empathy fatigue, participants with high empathy fatigue had lower perceived social support ( OR=0.406, 95% CI: 0.182-0.909, P<0.05). Except supporting primary and secondary traumatic stress, the total score of perceived social support and its dimensions were positively correlated with empathy satisfaction; and negatively correlated with job burnout and secondary traumatic stress. Regression analysis shows that empathy satisfaction had a positive predictive effect on overall scores of perceived social support and its dimensions of friend support and other support capacity. Conclusions:The social support status of community healthcare workers in Shanghai Minhang district is generally acceptable. However, it is important to pay special attention to the social support status of those who are under 40 years old, have a background in public health, are satisfied with their work, and experience a high level of empathy fatigue. Measures should be implemented to specifically enhance empathy satisfaction among healthcare workers; timely guidance and intervention should be provided accordingly.
7.Health economics evaluation of the community screening for chronic obstructive pulmonary disease based on family doctor contract services
Hongmei HUAN ; Shuai LIU ; Chi ZHANG ; Yonghong MU ; Weijin CAO ; Zhijun JIE ; Ming JIN
Chinese Journal of General Practitioners 2024;23(9):935-943
Objective:To explore the cost-effectiveness and cost-benefit of chronic obstructive pulmonary disease (COPD) screening and intervention based on family doctor contract services.Methods:From January 2023 to October 2023, using purposive sampling and a cross-sectional survey method, 1 040 individuals aged 60 to 75 years who underwent physical examinations at community health service centers in Shanghai Minhang district were selected as research subjects. The subjects were divided into a control group ( n=532) and a study group ( n=508). The control group received conventional community health education and participated in a COPD screening and intervention program provided by the examination center. The study group received COPD screening and intervention provided by contracted family doctors under a tightly integrated medical consortium. The screening content for both groups included initial and follow-up risk assessment questionnaires, pulmonary function tests, bronchodilator tests, and CT scans. A comparative analysis was conducted on the screening compliance rate, positive rate, and cost-effectiveness and cost-benefit between the two groups. Results:A total of 251 individuals at high risk for COPD and confirmed COPD patients were identified through initial and re-screening in both groups, accounting for 24.13% of the subjects, including 222 individuals at high risk (21.35%) and 29 confirmed patients (2.79%). The study group had significantly higher compliance rates in questionnaire-based initial screening, questionnaire-based re-screening+pulmonary function tests, bronchodilator experiments, diagnosis, and subsequent CT scans than the control group ( χ2=33.563, 41.425, 24.842, 17.363, all P<0.05).There were significant higher proportions of high-risk individuals and patients identified through screening in the study group than those in the control group ( χ2=44.880, all P<0.05). The study group had significantly higher positive rates of questionnaire-based initial screening, pulmonary function tests, bronchodilator experiments, and CT scans than the control group ( χ2=29.191, 11.313, 12.370, 4.429, all P<0.05). The overall costs of the screening in the control and study groups were 36 100 and 53 900 yuan, respectively. The cost of pulmonary function tests in the screening for the control and study groups was 1.44 million and 2.45 million yuan, respectively, accounting for 43.19% of the total cost, which was the highest proportion among all costs. The per capita cost for identifying one high-risk individual or patient with COPD was 358.46 Yuan, and the cost in study groups was significantly lower than that in the control group (322.75 vs. 429.45 yuan, χ2=20.396, P<0.05). The per capita net benefits and cost-benefit ratios for the overall subjects in the two groups were 0.37 million yuan and 43.50, respectively. The average net benefit of the study group and the control group was 0.49 million yuan and 0.26 million yuan respectively. The average net benefit of the study group was 0.23 million yuan higher than that of the control group. The cost-benefit ratio of the study group and the control group were 46.77 and 38.61, respectively. The cost-benefit ratio of the study group was 8.16 higher than that of the control group. Conclusion:The screening and intervention for COPD based on family doctor contract services have significant effectiveness, with a marked improvement in screening compliance and high cost-effectiveness, and can be used in community medical institutions with relatively sound family doctor and medical consortium services for targeted COPD screening.
8.Study on the Mechanism of Astragalosides Against Biliary Fibrosis by Inhibiting Ductular Reaction
Jing FANG ; Yonghong HU ; Yue LIANG ; Yongping MU ; Wei LIU ; Ping LIU ; Ying LYU ; Jiamei CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(7):2327-2335
Objective To study the partial mechanism of astragalosides(ASTs)against biliary fibrosis through inhibiting ductular reaction.Methods Rats were randomly divided into sham operation group,bile duct ligation(BDL)group and ASTs group(n=8 in each group).On the first day of the second week after BDL,the rats in ASTs group were given intragastric administration of ASTs for 3 weeks(160 mg·kg-1·d-1,once a day).Rats in sham operation group and BDL group were given the same volume of water.At the end of the fourth week,all rats were euthanasia.HE staining and sirius red staining were used to observe the pathological changes and collagen deposition.The degree of liver fibrosis was evaluated by semi-quantitative analysis of positive area of sirius red staining and the content of hydroxyproline.The expression changes of α smooth muscle actin(α-SMA),Desmin,cytokeratin(CK)19,CK7,epithelial cell adhesion molecule(Epcam),OV6 and lysyl oxidase(LOX)family proteins in liver tissue were detected by immunohistochemistry,Western blot,and real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).In vitro,hepatic progenitor cell line WB-F344 cells were induced by sodium butyrate to differentiate into biliary epithelial cells,intervented of ASTs,and collected after 4 days.The expression changes of CK19,LOXL1 and LOXL2 of cells were detected by qRT-PCR.Results Compared with BDL group,serum ALT and AST activities in ASTs group were significantly decreased(P<0.01).Histopathological injury of liver tissue was significantly reduced,Hyp content and percentage of positive area of sirius red were significantly decreased(P<0.01).Immunohistochemical staining showed that the positive expressions of α-SMA,Desmin,CK19,CK7,Epcam and OV6 were significantly decreased in the ASTs group.The mRNA expressions of α-SMA,CK7,LOX and LOXL1 were significantly decreased.The protein expressions of Epcam and LOXL1 were significantly reduced.In vitro results showed that the gene expressions of CK19,LOXL1 and LOXL2 were significantly increased after sodium butyrate induction(P<0.01).Compared with sodium butyrate group,the gene expressions of CK19,LOXL1 and LOXL2 were significantly decreased in ASTs group(P<0.01).Conclusion ASTs improved biliary fibrosis by inhibiting ductular reaction,and the mechanism may be related to the down-regulation of LOXL1.
9.Role of GDC-0449 in a rat model of liver fibrosis induced by carbon tetrachloride combined with 2-acetylaminofluorene
Yonghong HU ; Zhun XIAO ; Yadong FU ; Yue LIANG ; Linzhang ZHANG ; Wei LIU ; Yongping MU ; Chenghai LIU ; Ping LIU ; Jiamei CHEN
Journal of Clinical Hepatology 2022;38(2):334-341
Objective To investigate the intervention effect of GDC-0449, a hedgehog signaling pathway inhibitor, on rats with liver fibrosis induced by carbon tetrachloride (CCl 4 ) combined with 2-acetylaminofluorene (2-AAF). Methods A total of 18 female Fisher344 rats were randomly divided into normal group, CCl 4 /2-AAF group, and GDC-0449 group, with 6 rats in each group. The rats in the CCl 4 /2-AAF group and the GDC-0449 group were given subcutaneously injected 30% CCl 4 -olive oil solution at a dose of 2 mL/kg twice a week for 6 weeks to induce liver fibrosis; since week 7, in addition to the injection of CCl 4 -olive oil solution, the rats in these two groups were given 2-AAF (100 mg/kg/d) by gavage, and the rats in the GDC-0449 group were given GDC-0449 (25 mg/kg/d) by gavage, while those in the normal group were given an equal volume of olive oil solution by injection and normal saline by gavage. All rats were sacrificed at the end of week 9, and related samples were collected. HE staining and sirius red (SR) staining were used to observe the changes in liver histopathology and collagen deposition, and the semi-quantitative analysis of SR-positive area and Ishak score were used to evaluate fibrosis degree; the alkaline hydrolysis method was used to measure the level of hydroxyproline (Hyp) in liver tissue; immunohistochemistry, Western blot, and qRT-PCR were used to measure the expression of α-smooth muscle actin (α-SMA), type Ⅰ collagen (Col-Ⅰ), type Ⅳ collagen (Col-Ⅳ), cytokeratin 19 (CK19), cytokeratin 7 (CK7), the epithelial cell adhesion molecule Epcam, and the hedgehog signaling pathway in liver tissue; double immunofluorescence staining was used to observe the colocalization of CK19 and the oval cell marker OV6. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the normal group, the CCl 4 /2-AAF group had marked inflammatory cell aggregation and collagen deposition in liver tissue, with the formation of a pseudolobular structure, as well as significant increases in Hyp level and collagen positive area ratio in liver tissue ( P < 0.05), Ishak score ( P < 0.05), and the expression of α-SMA, Col-Ⅰ, Col-Ⅳ, Epcam, CK19, CK7, the transmembrane transporter Smoothened (Smo), Hedgehog ligand Desert Hedgehog (Dhh), the Indian Hedgehog membrane-binding receptor Patched (Ptch2), and glioma-related oncogenes Gli1, Gli2, and Gli3 (all P < 0.05); double immunofluorescence staining showed that CK19-positive cells also expressed OV6 in the liver tissue of rats in the CCl 4 /2-AAF group, with a significant increase compared with the normal group. Compared with the CCl 4 /2-AAF group, the GDC-0449 group had significant reductions in inflammatory cell aggregation and collagen deposition in liver tissue, Hyp level and collagen positive area ratio in liver tissue ( P < 0.05), Ishak score ( P < 0.05), and the expression of α-SMA, Epcam, CK19, CK7, Smo, Ptch2, Gli1, Gli2, and Gli3 (all P < 0.05); double immunofluorescence staining showed a significant reduction in the number of cells with co-expression of OV6 and CK19 in liver tissue. Conclusion The Hedgehog signaling pathway inhibitor GDC-0449 can significantly inhibit the progression of liver fibrosis induced by CCl 4 /2-AAF in rats, possibly by inhibiting hepatic stellate cell activation, collagen deposition, activation and proliferation of hepatic progenitor cells, and differentiation of hepatic progenitor cells into biliary epithelial cells.
10.Research advances in bone marrow mesenchymal stem cells and its exosomes in treatment of end-stage liver disease
Yue LIANG ; Yonghong HU ; Wei LIU ; Yongping MU ; Jiamei CHEN ; Ping LIU
Journal of Clinical Hepatology 2022;38(11):2643-2648
End-stage liver disease is the late stage of various acute and chronic liver diseases with high mortality and seriously threatens people's health. Liver transplantation is currently an effective treatment method for this disease, but its clinical application is greatly limited by the factors such as shortage of donors and high costs. In recent years, clinical and basic studies have shown that bone marrow mesenchymal stem cell and its exosomes have good clinical application prospects in the treatment of end-stage liver disease. This article reviews the mechanism of action and clinical application of bone marrow mesenchymal stem cell and its exosomes in the treatment of end-stage liver disease, so as to provide a reference for further research.

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