1.Effects of total flavonoids from Carthamus tinctorius L. on hepatic stellate cell activation based on miRNA-204/NUAK1/Hippo signaling axis
Mingqi LI ; Xiaolu ZHAO ; Chenlu ZHANG ; Yinghe WANG ; Yuehong MA
China Pharmacy 2026;37(3):311-316
OBJECTIVE To investigate the effects of total flavonoids from Carthamus tinctorius L. (TFCTL) on hepatic stellate cell (HSC) activation based on the microRNA (miRNA)-204/NUAK family SNF1-like kinase 1 (NUAK1)/Hippo signaling axis, thereby elucidating the potential mechanism underlying their antifibrotic effects. METHODS The HSC-T6 cells were divided into control group, model group, TFCTL low-concentration group (20 μg/mL), TFCTL medium-concentration group (40 μg/mL), and TFCTL high-concentration group (60 μg/mL). Except for control group, the remaining groups were treated with 5 ng/mL of transforming growth factor-β to induce the activation of hepatic stellate cells, followed by the addition of corresponding drug solutions/culture medium and incubation for 24 hours. Cell apoptosis was assessed, the expression levels of α-smooth muscle actin (α-SMA), type Ⅰ collagen (Collagen Ⅰ) and proteins associated with the Hippo/Yes-associated protein (YAP) pathway [YAP, large tumor suppressor kinase 1 (LATS1), and mammalian STE20-like kinase 1 (MST1)] were detected. Additionally, cell transfection was used to investigate the activity of the miRNA-204/NUAK1/Hippo signaling axis at both the genetic and protein levels. RESULTS After intervention with TFCTL, the apoptosis rate of HSC-T6 cells and the protein expressions of MST1 (except for the TFCTL high-concentration group) and LATS1 were significantly increased (P<0.05), while the protein expressions of α-SMA, CollagenⅠ, and YAP (except for the TFCTL medium-concentration group) were significantly decreased (P<0.05). Further results from cell transfection experiments revealed that after transfection with miRNA-204 mimics, the mRNA it’s protein expressions of α-SMA, CollagenⅠ, NUAK1, and YAP in HSC-T6 cells were significantly decreased (P<0.05), while the mRNA and protein expressions of LATS1 and the mRNA expression of MST1 were significantly increased (P<0.05). Conversely, the results were opposite following transfection with miRNA-204 inhibitors. CONCLUSIONS TFCTL can exert anti-hepatic fibrosis effects by up-regulating the expression of miRNA-204, thereby down- regulating the expressions of NUAK1, inactivating the Hippo/YAP pathway, which in turn suppresses the activation of HSC and promotes their apoptosis.
2.Clinical Study on Treatment of Non-alcoholic Steatohepatitis Patients with Dyslipidemia by Dizhuo Huayu Prescription with Catgut Embedding Therapy
Xiaoyan LIU ; Dongfang SHANG ; Lihui ZHANG ; Chenlu ZHAO ; Siying WANG ; Huaxin CHEN ; Wenxia ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):152-159
ObjectiveTo observe the clinical efficacy and safety of Dizhuo Huayu prescription combined with catgut embedding therapy in patients with nonalcoholic steatohepatitis (NASH) and dyslipidemia and explore the effect of the combined therapy on inflammatory cytokines interleukin (IL)-18 and IL-1β. MethodsA total of 82 patients with NASH and dyslipidemia from the Gastroenterology Department of the First Affiliated Hospital of Henan University of Chinese Medicine were randomly divided into a control group and a treatment group, with 41 patients in each group. The control group received Polyene Polyenylphosphatidylcholine Capsules, while the treatment group received Dizhuo Huayu prescription granules combined with catgut embedding. The treatment duration was 24 weeks for both groups. At weeks 0, 12, and 24, the traditional Chinese medicine (TCM) syndrome score, body mass index (BMI), liver fat content assessed by Fibroscan (CAP value), the level of alanine transaminase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and free fatty acid (FFA), and the expression of inflammatory cytokines IL-18 and IL-1β in serum were observed. Adverse reactions in both groups were recorded. ResultsA comparison of the comprehensive therapeutic effects between the two groups after 24 weeks of treatment revealed that the total effective rate was 62.16% (23/37) in the control group and 85.71% (30/35) in the treatment group, with a statistically significant difference (χ2 = 5.14, P<0.05). At weeks 12 and 24 after treatment, the TCM syndrome score, BMI, CAP value, TC, TG, LDL-C, and FFA were all significantly lower in both groups compared to pre-treatment levels, while the HDL-C level significantly increased (P<0.05). The effect was better at week 24 (P<0.05) than at week 12 (P<0.05), and the treatment group showed better outcomes than the control group at weeks 12 and 24 after treatment (P<0.05). After 24 weeks of treatment, both groups exhibited significant reductions in IL-18 and IL-1β levels (P<0.05). The treatment group demonstrated superior efficacy compared to the control group after treatment (P<0.05). Both groups experienced decreases in ALT, AST, and GGT levels after treatment (P<0.05). However, there were no statistically significant differences between the 12-week and 24-week post-treatment values within each group, nor were there significant differences between the two groups. No significant adverse reactions were observed in both groups. ConclusionThe Dizhuo Huayu prescription combined with catgut embedding therapy is safe and effective in treating patients with NASH and dyslipidemia, exhibiting hepatoprotective, anti-inflammatory, lipid-regulating, and weight-reducing effects.
3.Anti-inflammatory and anti-apoptotic effects and mechanism of total flavonoids of hawthorn leaves on rat intestinal epithelial cells
Kai WANG ; Pei LIU ; Kexin QI ; Jingyi WANG ; Chenlu SUN ; Danning SHI ; Hongyue CHEN ; Daoling HE ; Yan ZHU ; Ling GAN
Chinese Journal of Veterinary Science 2025;45(7):1450-1457
This study aims to investigate the anti-inflammatory and anti-apoptotic effects of total flavonoids of hawthorn leaves(TFHL)on lipopolysaccharide(LPS)-induced inflammatory injury in rat intestinal epithelial(IEC-6)cells,as well as the underlying mechanisms.An in vitro inflam-mation model was first established by treating IEC-6 cells with lipopolysaccharide(LPS).IEC-6 cells were then incubated with three concentrations of TFHL for 24 h prior to a further 24 h LPS treatment.RT-qPCR was used to quantify mRNA levels of the inflammatory genes COX-2 and iN-OS,while Western blotting was used to assess protein levels of the apoptotic markers Bax,cleaved Caspase-3,Bcl-2,and the JNK/p-JNK signaling pathway.Finally,cells were pretreated with TFHL and/or the JNK inhibitor SP600125 for 24 h before LPS exposure for 24 h,in order to evaluate the combined effects of TFHL and SP600125 on LPS-induced inflammatory cytokine expression and apoptotic protein levels in IEC-6 cells.The results showed that,compared with the LPS group,the mRNA level of COX-2 and iNOS in the 2.5,5.0,10.0 mg/L TFHL group and the Bax and Caspase-3 protein levels decreased significantly(P<0.01),and the Bcl-2 protein level was significantly higher(P<0.01),p-JNK protein level and p-JNK/JNK ratio decreased significantly(P<0.01);compared with the LPS group,the COX-2 and iNOS mRNA levels of the TFHL+LPS group de-creased significantly(P<0.01),Bax,and Caspase-3 protein levels decreased significantly(P<0.01),and the level of Bcl-2 protein increased significantly(P<0.05);compared with the LPS group,the COX-2 and iNOS mRNA levels of the TFHL+SP600125 group decreased significantly(P<0.01),Bax and Caspase-3 protein levels decreased significantly(P<0.01),and Bcl-2 protein level increased significantly(P<0.01).These findings indicate that TFHL exerts anti-inflammato-ry and anti-apoptotic effects in LPS-challenged IEC-6 cells by inhibiting the JNK signaling path-way.
4.Expression of serum hemoglobin β in hepatocellular carcinoma patients and its correlation with topoisomeraseⅡα
Jinhui TAN ; Xinxin XU ; Ping ZHAO ; Chenlu WANG ; Lining ZHOU ; Xiang CHEN ; Fuying CHU
Chinese Journal of Clinical Laboratory Science 2025;43(5):356-361
Objective To investigate the expression levels of serum hemoglobin β(HBB)in hepatocellular carcinoma(HCC)patients and its correlation with topoisomeraseⅡα(TOP2A)gene.Methods A total of 48 HCC patients visited the Second Affiliated Hospital of Nantong University from August 2023 to September 2024 were selected as the HCC group,and 32 healthy individuals who underwent physical examination during the same period were selected as the healthy control group.Their blood samples were collected,and the ex-pression levels of serum HBB and TOP2A genes were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Then,the relationship between the expression of HBB gene and clinicopathological parameters of the patients was ana-lyzed.The Kaplan-Meier Plotter database was used to evaluate the relationship between the expression of HBB gene and the prognosis of HCC.The diagnostic value of the expressions of serum HBB and TOP2A genes for HCC was assessed by the receiver operating charac-teristic(ROC)curve.Spearman rank correlation analysis was used to evaluate the correlation between the expressions of serum HBB and TOP2A genes in HCC patients.The regulatory effect of HBB gene on TOP2A gene was verified by the cell experiment.Results The expression levels of serum HBB gene in HCC patients(0.097[0.055,0.155])were significantly lower than that in healthy controls(1.029[0.625,1.434],U=19,P<0.001).The expression levels of serum TOP2A gene in HCC patients(1.810[0.825,3.623])were significantly higher than that in healthy controls(1.047[0.604,1.364],U=495,P=0.007).The expression level of serum HBB gene in HCC patients was significantly negatively correlated with that of TOP2A gene(ρ=-0.384,P=0.007).The analysis results of clinicopathological parameters showed that the expression level of HBB gene was only related to tumor size(χ2=4.090,P<0.05).The Kaplan-Meier survival analysis showed that the 5-year overall survival rate of the patients with low expression of HBB gene was signifi-cantly lower than that with high expression(HR=0.680,95%CI:0.470-0.970,P<0.05).The analysis of the ROC curve showed that the area under the ROC curve(AUCROC)of HBB gene in diagnosing HCC was 0.987(95%CI:0.963-1.000).When the cut-off value was 0.228,its sensitivity was 100%and specificity was 97%.The AUCROC of TOP2A gene for the diagnosis of HCC was 0.677(95%CI:0.559-0.797).When the cut-off value was 1.285,its sensitivity was 65%and specificity was 75%.The combined detection of HBB and TOP2A genes for the diagnosis of HCC had an AUCROC of 0.988(95%CI:0.965-1.000).When the cut-off value was 0.657,its sensitivity was 100%and specificity was 97%.The cell experiment results showed that the overexpression of HBB gene could inhibit the expression of TOP2A gene,while the knockout of HBB gene had the opposite effect.Conclusion HBB gene is lowly expressed in the serum of HCC patients and is significantly negatively correlated with the expression of TOP2A gene.
5.Validation of the effectiveness of leaf position data for the linear accelerator log file
Chenlu LIU ; Xiaotong WANG ; Long SUN ; Chong XU ; Bo XUE ; Xiaohua YANG ; Guoping SHAN
Chinese Journal of Radiation Oncology 2025;34(8):796-803
Objective:To validate the effectiveness of the leaf position data of multi-leaf collimator (MLC) recorded in the Infinity medical linear accelerator log file (LF).Methods:In this study, the establishment of the film dose calibration curve involved two steps. Initially, the pixel values obtained from scanning the film were converted into net optical density values. Subsequently, a sixth-order polynomial fit was performed on the net optical density and dose data. The picket fence (PF) tests were performed on the electronic portal imaging device (EPID) and film, and the corresponding acquired PF test images were analyzed using the relative leaf travel distance, relative leaf position, relative leaf pair full-width half -maximum (FWHM), and relative leaf pair neighbor region width analysis index, and compared with the relevant results in LF. In addition, to investigate the effect of gravity on the recorded leaf position data by the Infinity medical linear accelerator LF, the above PF tests were executed at 0°, 90°, 180°, and 270° gantry angles, respectively. Intergroup differences were assessed with the Wilcoxon rank-sum test.Results:The film dose calibration curve demonstrated high goodness-of-fit, with a correlation coefficient ( R2) of 0.9994. Across all gantry angles, the dispersion of overall leaf position data for EBT3 film, EPID, and LF followed a consistent pattern based on four analysis metrics: EBT3 film > EPID > LF. Moreover, for each analysis metric, inter-tool differences in standard deviations of results were consistently below 0.1 mm, and this pattern was invariant to gantry angle. Conclusions:The leaf position data recorded in the Infinity medical linear accelerator LF have a high degree of accuracy, and can be used as reference for the actual position of the MLC leaf. Furthermore, gravitational forces exhibit negligible impact on leaf position data acquired via LF.
6.Perioperative care of a patient undergoing total pancreatectomy with autogenous islet cell autotransplan-tation for pancreatic cancer
Chenlu HU ; Xiangru PEI ; Fang WANG ; Qian YU ; Yulian YING
Chinese Journal of Nursing 2025;60(7):880-883
To summarize the nursing experience of a patient with pancreatic cancer who underwent total pancreatectomy combined with islet cell transplantation.Nursing points:goal-oriented management of endocrine function,being alert to the occurrence of"brittle diabetes";the dose of pancreatic enzymes was dynamically adjusted by doctors and nurses to improve exocrine insufficiency;the whole-process nutritional risk assessment and individualized nutritional support strategies were implemented.Early identification and standardized management of gastroparesis syndrome can improve gastric motility of patients.After careful treatment and nursing care,the patient's islet function was stable,and the endocrine and exocrine functions of the pancreas recovered well.The patient was discharged 15 days after the operation.During the follow-up of 2 months,the patient's condition was stable,and no serious blood glucose disorder and other serious complications occurred.
7.Perioperative care of a patient undergoing total pancreatectomy with autogenous islet cell autotransplan-tation for pancreatic cancer
Chenlu HU ; Xiangru PEI ; Fang WANG ; Qian YU ; Yulian YING
Chinese Journal of Nursing 2025;60(7):880-883
To summarize the nursing experience of a patient with pancreatic cancer who underwent total pancreatectomy combined with islet cell transplantation.Nursing points:goal-oriented management of endocrine function,being alert to the occurrence of"brittle diabetes";the dose of pancreatic enzymes was dynamically adjusted by doctors and nurses to improve exocrine insufficiency;the whole-process nutritional risk assessment and individualized nutritional support strategies were implemented.Early identification and standardized management of gastroparesis syndrome can improve gastric motility of patients.After careful treatment and nursing care,the patient's islet function was stable,and the endocrine and exocrine functions of the pancreas recovered well.The patient was discharged 15 days after the operation.During the follow-up of 2 months,the patient's condition was stable,and no serious blood glucose disorder and other serious complications occurred.
8.Value of combined prediction of peripheral blood CD4+,CD3-CD16+CD56+,CD8+,CD19+for survival outcome in patients with EBV infection-associated hemophagocytic syndrome
Zhe PENG ; Pei XING ; Chenlu WANG ; Wenjie YU ; Yuli JIA
International Journal of Laboratory Medicine 2025;46(8):982-986,991
Objective To analyze the value of combined prediction of peripheral blood CD4+,CD3-CD16+CD56+,CD8+,CD19+for the risk of death in patients with Epstein-Barr virus-associated hemophagocytic syn-drome(EBV-HLH).Methods A total of 84 patients with EBV-HLH who were treated in Beijing Friendship Hospital Affiliated to Capital Medical University and Peking University First Hospital from May 2021 to A-pril 2023 were selected and divided into a death group(17 cases)and a survival group(67 cases)according to their survival outcomes within 6 months.The general information and peripheral blood T lymphocyte subsets of the two groups were compared,multivariate Logistic regression analysis was conducted to investigate the influencing factors of death in EBV-HLH patients,and receiver operating characteristic(ROC)curve was used to evaluate the predictive value of peripheral blood T lymphocyte subsets on death in EBV-HLH patients.Results The platelet count,hemoglobin,CD4+and CD3-CD16+CD56+levels in the death group were lower than those in the survival group,while the levels of CD8+and CD19+were higher than those in the survival group(P<0.05).Multivariate Logistic regression analysis showed that platelet count,hemoglobin,CD4+,and CD3-CD16+CD56+were all independent protective factors for the death of EBV-HLH patients,while CD8+and CD19+were independent risk factors for the death of EBV-HLH patients(P<0.05).The ROC curve results showed that the area under the curve of combined prediction of CD4+,CD8+,CD19+,CD3 CD16+CD56+for death in EBV-HLH patients was 0.923,with sensitivity and specificity of 94.12%and 80.60%,respectively,which were significantly higher than those predicted by each individual indicator(P<0.05).Conclusion The abnormal expression of CD4+,CD3-CD16+CD56+,CD8+,and CD19+in peripheral blood of EBV-HLH patients is associated with their survival outcomes,and the combined prediction of CD4+,CD3-CD16+CD56+,CD8+,and CD19+has high reference value for predicting patients'death.
9.Expression of serum hemoglobin β in hepatocellular carcinoma patients and its correlation with topoisomeraseⅡα
Jinhui TAN ; Xinxin XU ; Ping ZHAO ; Chenlu WANG ; Lining ZHOU ; Xiang CHEN ; Fuying CHU
Chinese Journal of Clinical Laboratory Science 2025;43(5):356-361
Objective To investigate the expression levels of serum hemoglobin β(HBB)in hepatocellular carcinoma(HCC)patients and its correlation with topoisomeraseⅡα(TOP2A)gene.Methods A total of 48 HCC patients visited the Second Affiliated Hospital of Nantong University from August 2023 to September 2024 were selected as the HCC group,and 32 healthy individuals who underwent physical examination during the same period were selected as the healthy control group.Their blood samples were collected,and the ex-pression levels of serum HBB and TOP2A genes were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Then,the relationship between the expression of HBB gene and clinicopathological parameters of the patients was ana-lyzed.The Kaplan-Meier Plotter database was used to evaluate the relationship between the expression of HBB gene and the prognosis of HCC.The diagnostic value of the expressions of serum HBB and TOP2A genes for HCC was assessed by the receiver operating charac-teristic(ROC)curve.Spearman rank correlation analysis was used to evaluate the correlation between the expressions of serum HBB and TOP2A genes in HCC patients.The regulatory effect of HBB gene on TOP2A gene was verified by the cell experiment.Results The expression levels of serum HBB gene in HCC patients(0.097[0.055,0.155])were significantly lower than that in healthy controls(1.029[0.625,1.434],U=19,P<0.001).The expression levels of serum TOP2A gene in HCC patients(1.810[0.825,3.623])were significantly higher than that in healthy controls(1.047[0.604,1.364],U=495,P=0.007).The expression level of serum HBB gene in HCC patients was significantly negatively correlated with that of TOP2A gene(ρ=-0.384,P=0.007).The analysis results of clinicopathological parameters showed that the expression level of HBB gene was only related to tumor size(χ2=4.090,P<0.05).The Kaplan-Meier survival analysis showed that the 5-year overall survival rate of the patients with low expression of HBB gene was signifi-cantly lower than that with high expression(HR=0.680,95%CI:0.470-0.970,P<0.05).The analysis of the ROC curve showed that the area under the ROC curve(AUCROC)of HBB gene in diagnosing HCC was 0.987(95%CI:0.963-1.000).When the cut-off value was 0.228,its sensitivity was 100%and specificity was 97%.The AUCROC of TOP2A gene for the diagnosis of HCC was 0.677(95%CI:0.559-0.797).When the cut-off value was 1.285,its sensitivity was 65%and specificity was 75%.The combined detection of HBB and TOP2A genes for the diagnosis of HCC had an AUCROC of 0.988(95%CI:0.965-1.000).When the cut-off value was 0.657,its sensitivity was 100%and specificity was 97%.The cell experiment results showed that the overexpression of HBB gene could inhibit the expression of TOP2A gene,while the knockout of HBB gene had the opposite effect.Conclusion HBB gene is lowly expressed in the serum of HCC patients and is significantly negatively correlated with the expression of TOP2A gene.
10.Validation of the effectiveness of leaf position data for the linear accelerator log file
Chenlu LIU ; Xiaotong WANG ; Long SUN ; Chong XU ; Bo XUE ; Xiaohua YANG ; Guoping SHAN
Chinese Journal of Radiation Oncology 2025;34(8):796-803
Objective:To validate the effectiveness of the leaf position data of multi-leaf collimator (MLC) recorded in the Infinity medical linear accelerator log file (LF).Methods:In this study, the establishment of the film dose calibration curve involved two steps. Initially, the pixel values obtained from scanning the film were converted into net optical density values. Subsequently, a sixth-order polynomial fit was performed on the net optical density and dose data. The picket fence (PF) tests were performed on the electronic portal imaging device (EPID) and film, and the corresponding acquired PF test images were analyzed using the relative leaf travel distance, relative leaf position, relative leaf pair full-width half -maximum (FWHM), and relative leaf pair neighbor region width analysis index, and compared with the relevant results in LF. In addition, to investigate the effect of gravity on the recorded leaf position data by the Infinity medical linear accelerator LF, the above PF tests were executed at 0°, 90°, 180°, and 270° gantry angles, respectively. Intergroup differences were assessed with the Wilcoxon rank-sum test.Results:The film dose calibration curve demonstrated high goodness-of-fit, with a correlation coefficient ( R2) of 0.9994. Across all gantry angles, the dispersion of overall leaf position data for EBT3 film, EPID, and LF followed a consistent pattern based on four analysis metrics: EBT3 film > EPID > LF. Moreover, for each analysis metric, inter-tool differences in standard deviations of results were consistently below 0.1 mm, and this pattern was invariant to gantry angle. Conclusions:The leaf position data recorded in the Infinity medical linear accelerator LF have a high degree of accuracy, and can be used as reference for the actual position of the MLC leaf. Furthermore, gravitational forces exhibit negligible impact on leaf position data acquired via LF.

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