1.Carvedilol to prevent hepatic decompensation of cirrhosis in patients with clinically significant portal hypertension stratified by new non-invasive model (CHESS2306)
Chuan LIU ; Hong YOU ; Qing-Lei ZENG ; Yu Jun WONG ; Bingqiong WANG ; Ivica GRGUREVIC ; Chenghai LIU ; Hyung Joon YIM ; Wei GOU ; Bingtian DONG ; Shenghong JU ; Yanan GUO ; Qian YU ; Masashi HIROOKA ; Hirayuki ENOMOTO ; Amr Shaaban HANAFY ; Zhujun CAO ; Xiemin DONG ; Jing LV ; Tae Hyung KIM ; Yohei KOIZUMI ; Yoichi HIASA ; Takashi NISHIMURA ; Hiroko IIJIMA ; Chuanjun XU ; Erhei DAI ; Xiaoling LAN ; Changxiang LAI ; Shirong LIU ; Fang WANG ; Ying GUO ; Jiaojian LV ; Liting ZHANG ; Yuqing WANG ; Qing XIE ; Chuxiao SHAO ; Zhensheng LIU ; Federico RAVAIOLI ; Antonio COLECCHIA ; Jie LI ; Gao-Jun TENG ; Xiaolong QI
Clinical and Molecular Hepatology 2025;31(1):105-118
Background:
s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model.
Methods:
Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort.
Results:
In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM).
Conclusions
Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.
2.Serum levels of trefoil factor 1 and bone morphogenetic protein 4 in patients with diabetic retinopathy and their clinical significance
Laixia DING ; Hongjuan XU ; Yunyi GU ; Yuzhe LIU ; Fang QIAN
International Eye Science 2025;25(7):1135-1139
AIM: To investigate the changes in serum levels of trefoil factor 1(Tff1)and bone morphogenetic protein 4(BMP4)in patients with diabetic retinopathy, and to evaluate their diagnostic value for diabetic retinopathy.METHODS: From January 2022 to January 2024, 186 patients with type 2 diabetes were selected as the study group and divided into a retinopathy subgroup(52 cases)and a non-retinopathy subgroup(134 cases)based on the presence of retinopathy. Another 186 healthy volunteers who underwent physical examinations during the same period were chosen as the control group. Serum Tff1 and BMP4 levels were measured using the enzyme-linked immunosorbent assay(ELISA). Pearson analysis was used to assess the correlation between serum Tff1, BMP4 levels, and clinical data. Multivariate Logistic regression analysis was performed to identify factors influencing the development of retinopathy in type 2 diabetic patients. Receiver operating characteristic(ROC)curve analysis was conducted to evaluate the diagnostic value of serum Tff1 and BMP4 levels for retinopathy in type 2 diabetic patients.RESULTS: Compared to the control group, serum Tff1 levels were lower and BMP4 levels were higher in both retinopathy and non-retinopathy subgroups(all P<0.05). Specifically, serum Tff1 levels were lower and BMP4 levels were higher in the retinopathy subgroup than in the non-retinopathy subgroup(all P<0.05). Pearson analysis revealed that Tff1 levels in type 2 diabetes patients were negatively correlated with disease duration, glycated hemoglobin levels, and triglyceride levels, while BMP4 levels were positively correlated(all P<0.05). Multivariate Logistic regression analysis identified type 2 diabetes duration, glycated hemoglobin, triglycerides, Tff1, and BMP4 as influencing factors for retinopathy development in type 2 diabetes patients(all P<0.05). ROC curve analysis showed that the combined diagnosis of serum Tff1 and BMP4 had an area under the curve(AUC)of 0.901, which was significantly higher than that of Tff1 alone(Z=2.069, P=0.039)and BMP4 alone(Z=2.072, P=0.038).CONCLUSION: Serum Tff1 levels are decreased and BMP4 levels are increased in patients with diabetic retinopathy, and the combined detection of these two markers offers high diagnostic value for diabetic retinopathy.
3.Analysis of depressive symptoms and associated factors among primary and secondary school students in the in depth monitoring counties Rural Nutrition Improvement Program
Chinese Journal of School Health 2025;46(2):219-222
Objective:
To understand the prevalence and related factors of depressive symptoms among primary and secondary school students in the in depth monitoring counties of China s Rural Compulsory Education Nutrition Improvement Program, so as to provide a basis for prevention and psychological intervention of depressive symptoms among children and adolescents in rural areas.
Methods:
In November 2022, a stratified random sampling method was adopted to collect height and weight data, basic personal and family information of 7 949 primary and secondary school students from grade three to grade nine through physical measurements and questionnaires in 56 key monitoring schools implementing the Student Nutrition Improvement Program in 7 in depth monitoring counties (Jalaid Banner in Inner Mongolia, Jinzhai County in Anhui, Mao Xian in Sichuan, Tiandeng County in Guangxi, Mian County in Shaanxi, Zhaozhou County in Heilongjiang and Youxi County in Fujian), and to obtain the information related to their depressive symptoms through the self assessment questionnaire on depression. Multivariate Logistic regression analysis was conducted to analyze the prevalence of depressive symptoms among primary and secondary school students, as well as their related factors.
Results:
The detection rate of depressive symptoms among primary and secondary school students in the in depth monitored counties was 23.5%. Logistic regression analysis showed that the probability of detecting depressive symptoms was higher among female students, middle school students, students whose video screen duration per day was >2 h, and students whose parents marital status was divorced or widowed ( OR =1.40, 1.64, 1.60, 1.24), and students whose sleep duration reached the recommended standard, whose parents usually accompanied them daily for time was 60-<120 min and ≥120 min, and students whose mothers literacy level was middle school graduation had lower probability of detecting depressive symptoms ( OR =0.85, 0.84, 0.71, 0.76) ( P < 0.05 ).
Conclusion
The detection rate of depressive symptoms among students in the in depth monitoring area is high, and targeted interventions need to be developed for students to reduce the risk of mental health problems.
4.Relationship between screening myopia and physical fitness index in college freshmen without majoring in public safety administration
Chinese Journal of School Health 2025;46(3):431-434
Objective:
To explore the relationship between visual acuity and physical fitness of university freshmen, so as to provide reference for myopia prevention and control for freshmen.
Methods:
From October to November 2022, 2 160 college freshman without majoring in public safety administration, selected from Guangxi Police College in 2022 by using the stratified cluster random sampling method, were reviewed for the results of visual acuity test and physical fitness scores. The physical fitness indices were evaluated by using the Z scores of physical fitness test scores, and the strength of association between the level of physical fitness index and myopia was analyzed by using Logistic regression model.
Results:
Among 2 160 college freshman without majoring in public safety administration, 917 (42.5%) students were diagnosed screening myopia, including 66 (3.1%) cases of high myopia, 383 (17.7%) cases of moderate myopia and 468 (21.7%) cases of mild myopia. The differences in the distribution of visual acuity tests among students with different physical fitness indices, body mass index, and gender were statistically significant ( Z/H=54.50, 49.53, 15.51, P <0.01). Low level and low middle level physical fitness indices were associated with screening myopia among freshmen[ OR (95% CI )=2.81(1.93-4.08),1.87(1.38-2.54)], and low level physical fitness indexes were associated with high myopia [ OR (95% CI )=7.22(2.33-22.32)] ( P <0.01).
Conclusions
Screening myopia among college freshman without majoring in public safety administration is related to physical fitness, and low level and low middle level physical fitness index are risk factors for myopia. Improving the level of physical fitness might be effective in preventing myopia.
5.Efficacy and Mechanism of Shuanghua Drink in Treating Primary Dysmenorrhea Based on COX-2/NF-κB Signaling Pathway
Yuncheng MA ; Yuanyuan SHI ; Zhen LIU ; Yuxi WANG ; Yuan TIAN ; Qian LI ; Xiaozhu WANG ; Cheng HE ; Wenhui XU ; Weiling WANG ; Jian GAO ; Ting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):72-80
ObjectiveTo evaluate the efficacy of Shuanghua drink in treating primary dysmenorrhea in the rat model and explore its mechanism of action. MethodsAn oxytocin-induced writhing mouse model was established to evaluate the analgesic effect of Shuanghua drink. Forty-eight non-pregnant female institute of cancer research (ICR) mice were randomly divided into six groups, including a blank group, a model group, an ibuprofen group (85.00 mg·kg-1), a low-dose group of Shuanghua drink (7.14 mL·kg-1), a medium-dose group of Shuanghua drink (14.28 mL·kg-1), and a high-dose group of Shuanghua drink (28.57 mL·kg-1). Each group consisted of eight mice. All treatment groups received daily intragastric administration at corresponding doses for 10 consecutive days. One hour after the final administration, 2 U of oxytocin was intraperitoneally injected per mouse. The writhing latency and number of writhing within 20 minutes were recorded. A primary dysmenorrhea rat model was established by using estradiol benzoate and oxytocin to evaluate the inhibitory effect of Shuanghua drink on the contraction of uterine smooth muscle. Forty-eight non-pregnant female Sprague-Dawley (SD) rats were divided into six groups, including a blank group, a model group, an ibuprofen group (51.00 mg·kg-1), a low-dose group of Shuanghua drink (4.28 mL·kg-1), a medium-dose group of Shuanghua drink (8.57 mL·kg-1), and a high-dose group of Shuanghua drink (17.10 mL·kg-1). Each group consisted of eight rats. Rats received subcutaneous injections of estradiol benzoate for 10 consecutive days to enhance uterine sensitivity. On the eleventh day, oxytocin (2 U/rat) was intraperitoneally administered to induce abnormal uterine contractions for establishing the primary dysmenorrhea model. All treatment groups received daily intragastric administration from the second day of modeling for 10 days. The effects of Shuanghua drink were evaluated by using parameters including uterine motility and the variation rate of uterine motility. The mechanism of action was investigated in rats with primary dysmenorrhea. The content of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), thromboxane B2 (TXB2), prostacyclin metabolite (6-keto-PGF1α), and β-endorphin (β-EP) in uterine tissue of rats was detected by using enzyme-linked immunosorbent assay (ELISA). The changes in the content of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) were analyzed via colorimetric assay. Western blot was performed to determine the content of phosphorylated inhibitor of kappa B kinase beta (p-IKKβ)/IKKβ, phosphorylated inhibitor of kappa B alpha (p-IκBα), IκBα, phosphorylated p65 (p-p65), p65, and cyclooxygenase-2 (COX-2) proteins in uterine tissue of rats. ResultsIn the oxytocin-induced writhing mouse model, the model group exhibited significantly shortened writhing latency and increased writhing frequency compared to the control group (P<0.01). Both the ibuprofen group and the high-dose group of Shuanghua drink displayed prolonged writhing latency (P<0.05), while the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink exhibited reduced writhing frequency (P<0.01). In the primary dysmenorrhea rat model, the uterine motility and its variation rate in the model group were significantly higher than those in the blank group (P<0.01). These parameters were markedly suppressed by ibuprofen and Shuanghua drink at all tested doses (P<0.01). For the mechanism of action, the model group showed significantly increased PGF2α/PGE2, TXB2/6-keto-PGF1α, NO, and iNOS in uterine tissue (P<0.05, P<0.01) and significantly decreased β-EP (P<0.01). These parameters were significantly attenuated in the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink. The PGF2α/PGE2 (P<0.01), TXB2/6-keto-PGF1α (P<0.01), NO (medium-dose group P<0.05), and iNOS (P<0.01) were reduced, and the β-EP (medium-dose group P<0.05) was up-regulated. Compared to the model group, the ibuprofen group and medium-dose group of Shuanghua drink showed significantly increased content of β-EP in the serum of rats (P<0.05). Compared to the blank group, the model group showed significantly elevated expressions of COX-2, p-IKKβ/IKKβ, p-IκBα/IκBα, and p-p65/p65 proteins (P<0.01) and significantly reduced anti-inflammatory protein IκBα (P<0.05). Compared to the model group, the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink showed significantly reduced expressions of COX-2 (P<0.01), p-IKKβ/IKKβ (P<0.01), p-IκBα/IκBα (P<0.05, P<0.01), and p-p65/p65(P<0.01) and up-regulated expression of IκBα protein (P<0.05, P<0.01). ConclusionShuanghua drink effectively alleviates primary dysmenorrhea through analgesia and suppression of abnormal contractions of uterine smooth muscle. Its mechanism may be mediated by reduced levels of PGF2α/PGE2, TXB2/6-keto-PGF1α, iNOS, and NO, elevated β-EP level, and inhibited COX-2/NF-κB signaling pathway.
6.Efficacy and Mechanism of Shuanghua Drink in Treating Primary Dysmenorrhea Based on COX-2/NF-κB Signaling Pathway
Yuncheng MA ; Yuanyuan SHI ; Zhen LIU ; Yuxi WANG ; Yuan TIAN ; Qian LI ; Xiaozhu WANG ; Cheng HE ; Wenhui XU ; Weiling WANG ; Jian GAO ; Ting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):72-80
ObjectiveTo evaluate the efficacy of Shuanghua drink in treating primary dysmenorrhea in the rat model and explore its mechanism of action. MethodsAn oxytocin-induced writhing mouse model was established to evaluate the analgesic effect of Shuanghua drink. Forty-eight non-pregnant female institute of cancer research (ICR) mice were randomly divided into six groups, including a blank group, a model group, an ibuprofen group (85.00 mg·kg-1), a low-dose group of Shuanghua drink (7.14 mL·kg-1), a medium-dose group of Shuanghua drink (14.28 mL·kg-1), and a high-dose group of Shuanghua drink (28.57 mL·kg-1). Each group consisted of eight mice. All treatment groups received daily intragastric administration at corresponding doses for 10 consecutive days. One hour after the final administration, 2 U of oxytocin was intraperitoneally injected per mouse. The writhing latency and number of writhing within 20 minutes were recorded. A primary dysmenorrhea rat model was established by using estradiol benzoate and oxytocin to evaluate the inhibitory effect of Shuanghua drink on the contraction of uterine smooth muscle. Forty-eight non-pregnant female Sprague-Dawley (SD) rats were divided into six groups, including a blank group, a model group, an ibuprofen group (51.00 mg·kg-1), a low-dose group of Shuanghua drink (4.28 mL·kg-1), a medium-dose group of Shuanghua drink (8.57 mL·kg-1), and a high-dose group of Shuanghua drink (17.10 mL·kg-1). Each group consisted of eight rats. Rats received subcutaneous injections of estradiol benzoate for 10 consecutive days to enhance uterine sensitivity. On the eleventh day, oxytocin (2 U/rat) was intraperitoneally administered to induce abnormal uterine contractions for establishing the primary dysmenorrhea model. All treatment groups received daily intragastric administration from the second day of modeling for 10 days. The effects of Shuanghua drink were evaluated by using parameters including uterine motility and the variation rate of uterine motility. The mechanism of action was investigated in rats with primary dysmenorrhea. The content of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), thromboxane B2 (TXB2), prostacyclin metabolite (6-keto-PGF1α), and β-endorphin (β-EP) in uterine tissue of rats was detected by using enzyme-linked immunosorbent assay (ELISA). The changes in the content of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) were analyzed via colorimetric assay. Western blot was performed to determine the content of phosphorylated inhibitor of kappa B kinase beta (p-IKKβ)/IKKβ, phosphorylated inhibitor of kappa B alpha (p-IκBα), IκBα, phosphorylated p65 (p-p65), p65, and cyclooxygenase-2 (COX-2) proteins in uterine tissue of rats. ResultsIn the oxytocin-induced writhing mouse model, the model group exhibited significantly shortened writhing latency and increased writhing frequency compared to the control group (P<0.01). Both the ibuprofen group and the high-dose group of Shuanghua drink displayed prolonged writhing latency (P<0.05), while the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink exhibited reduced writhing frequency (P<0.01). In the primary dysmenorrhea rat model, the uterine motility and its variation rate in the model group were significantly higher than those in the blank group (P<0.01). These parameters were markedly suppressed by ibuprofen and Shuanghua drink at all tested doses (P<0.01). For the mechanism of action, the model group showed significantly increased PGF2α/PGE2, TXB2/6-keto-PGF1α, NO, and iNOS in uterine tissue (P<0.05, P<0.01) and significantly decreased β-EP (P<0.01). These parameters were significantly attenuated in the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink. The PGF2α/PGE2 (P<0.01), TXB2/6-keto-PGF1α (P<0.01), NO (medium-dose group P<0.05), and iNOS (P<0.01) were reduced, and the β-EP (medium-dose group P<0.05) was up-regulated. Compared to the model group, the ibuprofen group and medium-dose group of Shuanghua drink showed significantly increased content of β-EP in the serum of rats (P<0.05). Compared to the blank group, the model group showed significantly elevated expressions of COX-2, p-IKKβ/IKKβ, p-IκBα/IκBα, and p-p65/p65 proteins (P<0.01) and significantly reduced anti-inflammatory protein IκBα (P<0.05). Compared to the model group, the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink showed significantly reduced expressions of COX-2 (P<0.01), p-IKKβ/IKKβ (P<0.01), p-IκBα/IκBα (P<0.05, P<0.01), and p-p65/p65(P<0.01) and up-regulated expression of IκBα protein (P<0.05, P<0.01). ConclusionShuanghua drink effectively alleviates primary dysmenorrhea through analgesia and suppression of abnormal contractions of uterine smooth muscle. Its mechanism may be mediated by reduced levels of PGF2α/PGE2, TXB2/6-keto-PGF1α, iNOS, and NO, elevated β-EP level, and inhibited COX-2/NF-κB signaling pathway.
7.Carvedilol to prevent hepatic decompensation of cirrhosis in patients with clinically significant portal hypertension stratified by new non-invasive model (CHESS2306)
Chuan LIU ; Hong YOU ; Qing-Lei ZENG ; Yu Jun WONG ; Bingqiong WANG ; Ivica GRGUREVIC ; Chenghai LIU ; Hyung Joon YIM ; Wei GOU ; Bingtian DONG ; Shenghong JU ; Yanan GUO ; Qian YU ; Masashi HIROOKA ; Hirayuki ENOMOTO ; Amr Shaaban HANAFY ; Zhujun CAO ; Xiemin DONG ; Jing LV ; Tae Hyung KIM ; Yohei KOIZUMI ; Yoichi HIASA ; Takashi NISHIMURA ; Hiroko IIJIMA ; Chuanjun XU ; Erhei DAI ; Xiaoling LAN ; Changxiang LAI ; Shirong LIU ; Fang WANG ; Ying GUO ; Jiaojian LV ; Liting ZHANG ; Yuqing WANG ; Qing XIE ; Chuxiao SHAO ; Zhensheng LIU ; Federico RAVAIOLI ; Antonio COLECCHIA ; Jie LI ; Gao-Jun TENG ; Xiaolong QI
Clinical and Molecular Hepatology 2025;31(1):105-118
Background:
s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model.
Methods:
Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort.
Results:
In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM).
Conclusions
Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.
8.Isolation and protection of organs at risk by crosslinked sodium hyaluronate gel during brachytherapy
Jianjian LIU ; Yan ZHANG ; Zhiwei CUI ; Dongfang WANG ; Xu LIU ; Shenglin YANG ; Qian CHAI ; Fenglin LIU
Chinese Journal of Tissue Engineering Research 2025;29(4):700-706
BACKGROUND:Crosslinked sodium hyaluronate gel has good mechanical property,biocompatibility,and biodegradability,and can be used as an isolated protective material in tumor radiation therapy to protect endangered organs from damage caused by excess radiation dose. OBJECTIVE:To investigate the safety and efficacy of crosslinked sodium hyaluronate gel in reducing the dose of radiation to dangerous organs during brachytherapy. METHODS:A total of 16 specific pathogen-free Kunming mice of the same age and similar body weight were selected as experimental subjects and divided into experimental group and control group by the random number table method,with 8 mice in each group.125I seeds were implanted subcutaneously in the back of mice in the experimental group,and then crosslinked sodium hyaluronate gel was injected around the radioactive particles.Only 125I seeds were implanted subcutaneously in the back of mice in the control group.After injection,the distance between the radioactive particles and the epidermis was measured by spiral CT scan,and the surface radiation dose was measured by radiation dosimeter.Within 10 weeks after injection,the growth state,survival rate,skin radiation damage,and gel retention of mice were observed. RESULTS AND CONCLUSION:(1)Spiral CT scan showed that the implanted gel was relatively concentrated and created an effective distance between the radioactive seeds and the epidermis.The body surface radiation dose of the experimental group was significantly lower than that of the control group(P<0.01).(2)During the experimental observation period,mice in both groups survived;mice in the control group showed obvious irritability and other unstable behavior in the late experimental period,and some mice in the experimental group showed similar behavior.The daily food intake of mice in the two groups had no significant change,and the body mass showed the same increasing trend.After implantation of radioactive seeds,the two groups of mice showed different degrees of radioactive skin injury.From day 23 after injection to the end of the experiment,the skin radiation injury score of the experimental group was lower than that of the control group(P<0.01).At week 10 after implantation,6 mice in the experimental group had no obvious gel residue under their skin,and 2 mice had a very small amount of scattered gel-like samples under their skin.(3)Therefore,the crosslinked sodium hyaluronate injection technique can increase the space between the radioactive target area of 125I seeds and the organ at risk outside the target through physical space occupying,which can effectively reduce the dose of the organ at risk,and play a role in the isolation and protection of the organ at risk.
9.Metformin exerts a protective effect on articular cartilage in osteoarthritis rats by inhibiting the PI3K/AKT/mTOR pathway
Tianjie XU ; Jiaxin FAN ; Xiaoling GUO ; Xiang JIA ; Xingwang ZHAO ; Kainan LIU ; Qian WANG
Chinese Journal of Tissue Engineering Research 2025;29(5):1003-1012
BACKGROUND:Studies have shown that metformin has anti-inflammatory,anti-tumor,anti-aging and vasoprotective effects,and can inhibit the progression of osteoarthritis,but its specific mechanism of action remains unclear. OBJECTIVE:To investigate the mechanism of metformin on cartilage protection in a rat model of osteoarthritis. METHODS:Forty male Sprague-Dawley rats were randomly divided into four groups(n=10 per group):blank,control,sham-operated,and metformin groups.The blank group did not undergo any surgery.In the sham-operated group,the joint cavity was exposed.In the model group and the metformin group,the modified Hulth method was used to establish the osteoarthritis model.At 1 day after modeling,the rats in the metformin group were given 200 mg/kg/d metformin by gavage,and the model,blank,and sham-operated groups were given normal saline by gavage.Administration in each group was given for 4 weeks consecutively.Hematoxylin-eosin staining,toluidine blue staining,and safranin O-fast green staining were used to observe the morphological structure of rat knee joints.Immunohistochemical staining and western blot were used to detect the protein expression of SOX9,type Ⅱ collagen,a disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTS5),Beclin1,P62,phosphatidylinositol 3-kinase(PI3K),p-PI3K,protein kinase B(AKT),p-AKT,mammalian target of rapamycin(Mtor),and p-Mtor in rat cartilage tissue. RESULTS AND CONCLUSION:The results of hematoxylin-eosin,toluidine blue and safranin O-fast green staining showed smooth cartilage surface of the knee joints and normal histomorphology in the blank group and the sham-operated group,while in the model group,there was irregular cartilage surface of the knee joint and cartilage damage,with a decrease in the number of chondrocytes and the content of proteoglycans in the cartilage matrix.In the metformin group,there was a significant improvement in the damage to the structure of the cartilage in the knee joints of the rats,and the cartilage surface tended to be smooth,with an increase in the number of chondrocytes and the content of proteoglycans in the cartilage matrix.Immunohistochemistry staining and western blot results showed that compared with the control and sham-operated groups,the expression of SOX9,type Ⅱ collagen,and Beclin1 proteins in the cartilage tissue of rats in the model group was significantly decreased(P<0.05).Conversely,the expression of ADAMTS5,P62,as well as p-PI3K,p-AKT,and p-Mtor proteins was significantly increased(P<0.05).Furthermore,compared with the model group,the expression of SOX9,type Ⅱ collagen,and Beclin1 proteins in the cartilage tissue of rats in the metformin group was significantly increased(P<0.05),while the expression of ADAMTS5,P62,as well as p-PI3K,p-AKT,and p-Mtor proteins was significantly decreased(P<0.05).To conclude,Metformin can improve the autophagy activity of chondrocytes and reduce the degradation of cartilage matrix in osteoarthritis rats by inhibiting the activation of PI3K/AKT/Mtor signaling pathway,thus exerting a protective effect on articular cartilage.
10.Compound Xishu Granules Inhibit Proliferation of Hepatocellular Carcinoma Cells by Regulating Ferroptosis
Yuan TIAN ; Yuxi WANG ; Zhen LIU ; Yuncheng MA ; Hongyu ZHU ; Xiaozhu WANG ; Qian LI ; Jian GAO ; Weiling WANG ; Wenhui XU ; Ting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):37-45
ObjectiveTo study the mechanism of compound Xishu granules (CXG) in inhibiting the proliferation of hepatocellular carcinoma cells by regulating ferroptosis. MethodsThe transplanted tumor model of human Huh7 was established with nude mice and the successfully modeled mice were randomized into model, Fufang Banmao (0.21 g·kg-1), low-dose (1.87 g·kg-1) CXG, medium-dose (3.74 g·kg-1) CXG, and high-dose (7.49 g·kg-1) CXG groups. Mice were administrated with drinking water or CXG for 28 days, and the body weight and tumor volume were measured every 4 days. Hematoxylin-eosin staining was employed to observe the histopathological changes of tumors. The cell-counting kit-8 (CCK-8) was used to examine the survival rate of Huh7 cells treated with different concentrations (0, 31.25, 62.5, 125, 250, 500, 1 000 mg·L-1) of CXG for 24 h and 48 h. CA-AM, DCFH-DA, and C11-BODIPY581/591 fluorescent probes were used to determine the intracellular levels of ferrous ion (Fe2+), reactive oxygen species (ROS), and lipid peroxide (LPO), respectively. The colorimetric method was employed to measure the levels of glutathione (GSH) and superoxide dismutase (SOD). Western blot was employed to determine the protein levels of glutathione peroxidase 4 (GPX4), transferrin receptor 1 (TFR1), and ferritin heavy chain 1 (FTH1), respectively. ResultsIn the animal experiment, compared with the model group, the drug treatment groups showed reductions in the tumor volume from day 12 (P<0.01). After treatment, the Fufang Banmao and low-, medium-, and high-dose CXG groups had lower tumor volume, relative tumor volume, and tumor weight than the model group (P<0.05), with tumor inhibition rates of 48.99%, 79.93%, 91.38%, and 97.36%, respectively. Moreover, the CXG groups had lower tumor volume and relative tumor volume (P<0.05 in all the three dose groups) and lower tumor weight (P<0.05 in medium-dose and high-dose groups) than the Fufang Banmao group. Compared with the model group, the drug treatment groups showed reduced number of tumor cells, necrotic foci with karyopyknosis, nuclear fragmentation, and nucleolysis, and the high-dose CXG group showed an increase in the proportion of interstitial fibroblasts. In the cell experiment, compared with the blank group, CXG reduced the survival rate of Huh7 cells in a dose-dependent manner after incubation for 24 h and 48 h (P<0.05). Compared with the blank group, the RSL3 group and the low-, medium-, and high-dose CXG groups showed a decrease in the relative fluorescence intensity of CA-AM and increases in the fluorescence intensity of DCFH-DA and fluorescence ratio of C11-BODIPY581/591, which indicated elevations in the levels of Fe2+ (P<0.01), ROS (P<0.05), and LPO (P<0.01), respectively. Compared with the blank group, the RSL3 and low-, medium-, and high-dose CXG groups showed lowered levels of GSH and SOD (P<0.05). In addition, the RSL3 group and the medium- and high-dose CXG groups showed down-regulated expression of GPX4 and FTH1 (P<0.05), and the low- and high-dose CXG groups presented up-regulated expression of TFR1 (P<0.05). ConclusionCXG suppresses the proliferation of hepatocellular carcinoma cells by inducing ferroptosis via downregulating the GSH-GPX4 signaling axis and increasing intracellular Fe2+and LPO levels.


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