1.The Effect of Qishao Tongbi Capsule (芪芍通痹胶囊) on the Wnt/β-catenin Pathway in a Rat Model of Intervertebral Disc Degeneration
Yumen XUE ; Xilin XU ; Wei HAN ; Jiaben XU ; Wenting XU ; Zelin LIU ; Xiaofeng ZHANG
Journal of Traditional Chinese Medicine 2025;66(1):79-88
ObjectiveTo explore the possible mechanism of Qishao Tongbi Capsule (芪芍通痹胶囊, QTC) in the treatment of intervertebral disc degeneration (IDD). MethodsSeventy-five rats were randomly divided into control group, model group, low-dose QTC group, high-dose QTC group, high-dose QTC +agonist group, with 15 rats in each group. Except for the control group, all other groups were subjected to a fibrous ring puncture to prepare an IDD model. After modeling, rats in low-dose QTC group and high-dose QTC group were given QTC at doses of 0.2 and 0.8 g/(kg·d) by gavage, respectively. Rats in high-dose QTC+ agonist group was given QTC at 0.8 g/(kg·d) and SKL2001 solution at 10 mg/(kg·d) by gavage. The control group and model group were given 10 ml/(kg·d) distilled water by gavage. All treatments were given once a day for 4 consecutive weeks. After treatment, X-ray and magnetic resonance imaging (MRI) were used to detect IDD degree. Hematoxylin-eosin (HE) staining and Safranin O-Fast Green staining were used to observe the morphological changes of the intervertebral disc tissue. Immunohistochemical staining was performed to examine the levels of proteoglycan, type Ⅱ collagen (COL Ⅱ), and matrix metalloproteinase-3 (MMP-3) in the intervertebral disc tissue. Western blotting was used to detect the extracellular matrix (ECM)-related proteins (proteoglycan, COL Ⅱ, MMP-3, MMP-9, MMP-13), aging-related proteins (P53, P21, P16), apoptosis related proteins, including B-cell lymphoma/leukemia 2 (BCL-2), BCL-2 related X protein (BAX), Cleaved Caspase-3, and Wnt/β-catenin pathway related proteins such as Wnt3a, glycogen synthase kinase-3β (GSK-3β) and β-catenin in the intervertebral disc nucleus pulposus (NP) tissue. Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) was used to assess the mRNA expression of Wnt3a, GSK-3β, and β-catenin in intervertebral disc tissue. ResultsCompared with the model group, rats in the low-dose QTC group and high-dose QTC group exhibited improved DHI, decreased Pfirmann grading, and alleviated IDD. The structural integrity of the NP and annulus fibrosus increased, and the number of the NP increased. The levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β increased, while the levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX, Cleaved Caspase-3, Wnt3a and β-catenin protein decreased. The mRNA expression of Wnt3a and β-catenin mRNA decreased, while GSK-3β mRNA expression increased (P<0.05). Compared with the low-dose QTC group, the high-dose QTC group showed further improvements in DHI, decrease in Pfirrmann grading (P<0.05), and greater alleviation of IDD. The structural integrity of NP and annulus fibrosus was further enhanced, and the number of NP cells further increased. The levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β were higher, while the levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX, Cleaved Caspase-3, Wnt3a and β-catenin were lower. The mRNA expression of Wnt3a and β-catenin decreased, while GSK-3β mRNA expression increased (P<0.05). Compared with the high-dose QTC group, the high-dose QTC +agonist group showed a decrease of DHI, an increase of Pfirmann grading (P<0.05), significant aggravation of IDD, reduction in structural integrity of the NP and annulus fibrosus, a decrease of NP cell count, lower levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β, and higher levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX and Cleaved Caspase-3. Additionally, GSK-3β mRNA expression decreased (P<0.05). ConclusionQTC can inhibit NP cell aging, apoptosis, and ECM degradation in IDD rats, and its therapeutic effect may be mediated through the inhibition of the Wnt/β-catenin pathway.
2.Label-free electrochemical aptasensing of cardiac cell secretomes in cell culture media for the evaluation of drug-induced myocardial injury.
Zelin YANG ; Xilin CHEN ; Mingang LIAO ; Feng LIAO ; Wen CHEN ; Qian SHAO ; Bing LIU ; Duanping SUN
Journal of Pharmaceutical Analysis 2025;15(10):101234-101234
Cardiac troponin I (cTnI), a widely used biomarker for assessing cardiovascular risk, can provide a window for the evaluation of drug-induced myocardial injury. Label-free biosensors are promising candidates for detecting cell secretomes, since they do not require labor-intensive processes. In this work, a label-free electrochemical aptasensor is developed for in situ monitoring of cardiac cell secretomes in cell culture media based on target-induced strand displacement. The aptasensing system contains an aptamer-functionalized signal nanoprobe facing trimetallic metal-organic framework nanosheets and a gold nanoparticle-based detection working electrode modified with DNA nanotetrahedron-based complementary DNA for indirect target detection. The signal nanoprobes (termed CAHA) consisted of copper-based metal-organic frameworks, AuPt nanoparticles, horseradish peroxidase, and an aptamer. When the aptasensor is exposed to cardiac cell secretomes, cTnI competitively binds to the aptamer, resulting in the release of signal nanoprobes from the biorecognition interface and electrochemical signal changes. The aptasensor exhibited rapid response times, a low detection limit of 0.31 pg/mL, and a wide linear range of 0.001-100 ng/mL. We successfully used this aptasensor to measure cTnI concentrations among secreted cardiac markers during antitumor drug treatment. In general, aptasensors can be used to monitor a variety of cardiac biomarkers in the evaluation of cardiotoxicity.
3.Effect of Liangxue Tuizi Formula (凉血退紫方) on RAF/MEK/ERK Pathway in Skin Tissue and Serum NETs Biomarkers in Henoch-Schönlein Purpura Model Rats with Blood Heat Syndrome
Yingying JIANG ; Manxiang YANG ; Zhenhua YUAN ; Leying XI ; Mingyang CAI ; Diya MA ; Yifan LI ; Yuhang NIU ; Runze LIU ; Jiawen CAO ; Xilin CHEN ; Xianqing REN
Journal of Traditional Chinese Medicine 2025;66(23):2475-2483
ObjectiveTo investigate the potential mechanism of Liangxue Tuizi Formula (凉血退紫方, LXTZF) in treating Henoch-Schönlein Purpura (HSP) by examining its regulatory effect on neutrophil extracellular trap (NETs) dysregulation via the rapidly accelerated fibrosarcoma kinase (RAF)/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway. MethodsSeventy Wistar rats were randomly allocated into a blank control group (n=14) and a modeling group (n=56). Rats in the modelling group underwent an eight-week modelling period to establish HSP rat models with blood-heat syndrome via modified ovalbumin (OVA) induction method combined with oral administration of heat-property Chinese herbal medicine. Fifty successfully modeled rats were subsequently randomly divided into five groups (n=10 per group), model group, compound glycyrrhizin group, LXTZF group, RAF inhibitor group, and LXTZF + RAF agonist group. Additionally, 10 rats were selected from the original blank control group for the final experiment. From the 11th week of modelling, rats in the blank control group and the model group received 1 ml/(100 g·d) ultrapure water via oral administration, in addition to 0.5 ml/(kg·d) 0.9% sodium chloride solution via intraperitoneal injection. The LXTZF group and the compound glycyrrhizin group received 7.5 g/(kg·d) LXTZF granule suspension via gavage, 13.5 mg/(kg·d) compound glycyrrhizin suspension via gavage, respectively. The RAF inhibitor group received 1 mg/(kg·d) GW5074 suspension via intraperitoneal injection and ultrapure water via oral administration; the LXTZF + RAF agonist group received 7.5 g/(kg·d) LXTZF granule suspension via gavage and 1 mg/(kg·d) paclitaxel suspension via intraperitoneal injection. All administrations were performed once daily for 4 weeks. After intervention, skin tissue histopathology was examined by hematoxylin and eosin (H&E) staining, immunoglobulin A (IgA) deposition was assessed via immunofluorescence, serum levels of neutrophil elastase (NE), tumor necrosis factor-α (TNF-α), and vascular cell adhesion molecule-1 (VCAM-1) were measured using enzyme-linked immunosorbent assay (ELISA), serum myeloperoxidase (MPO) level was determined by a colorimetric assay; the mRNA expression levels of RAF, MEK, and ERK in skin tissue were detected by real-time quantitative polymerase chain reaction (RT-qPCR); and the protein expression of RAF, MEK, ERK, as well as phosphorylated MEK (p-MEK) and phosphorylated ERK (p-ERK), were analyzed by Western Blot. ResultsSkin tissue in the blank control group rats remained normal, whereas the model group exhibited neutrophil infiltration and haemorrhage with red blood cell rupture. In all drug intervention groups, neutrophil infiltration and haemorrhagic exudation reduced markedly, with LXTZF group demonstrating the most pronounced improvement. Compared with the blank control group, rats in the model group exhibited enhanced IgA fluorescence intensity in skin tissue, elevated serum levels of NE, MPO, TNF-α and VCAM-1, increased mRNA expression of RAF, MEK, ERK1 and ERK2, as well as heightened RAF protein levels and p-MEK/MEK and p-ERK/ERK ratios (P<0.05). Compared with the model group, the drug intervention groups exhibited reduced IgA fluorescence intensity in skin tissue, along with decreased serum levels of NE, MPO, TNF-α, and VCAM-1 (P<0.05). In LXTZF group and RAF inhibition groups, reduced mRNA expression of RAF, MEK, ERK1, and ERK2 was observed in rat skin tissue, alongside decreased RAF protein levels and reduced p-MEK/MEK and p-ERK/ERK ratios (P<0.05). Compared with LXTZF + RAF agonist group, the compound glycyrrhizin group, LXTZF group, and RAF inhibitior group exhibited reduced IgA fluorescence intensity in skin tissue, decreased serum NE, MPO, TNF-α, and VCAM-1 levels, and decreased MEK mRNA expression and p-MEK/MEK ratio (P<0.05). ConclusionThe potential mechanism by which LXTZF treats Henoch-Schönlein purpura with blood heat syndrome may involve blocking the RAF/MEK/ERK signaling pathway in skin tissue, and suppressing excessive formation of NETs, thereby reducing IgA deposition in dermal microvessels and attenuating systemic inflammatory responses.
4.Jianpi-Huayu decotion combined with gemcitabine induces ferroptosis and inhibits growth of pancreatic cancer cells through Nrf2/SLC7A11/GPX4 axis
Xinqiu CHEN ; Zhengze ZHANG ; Shuwei LIU ; Xiaoyu ZHU ; Yongheng LAI ; Chongkai FANG ; Junhai HUANG ; Xilin ZHAO ; Chong ZHONG
Chinese Journal of Pathophysiology 2025;41(6):1077-1087
AIM:To investigate the effect of Jianpi-Huayu decoction(JPHYD)combined with gemcitabine(GEM)on ferroptosis of pancreatic cancer PANC-1 cells and its mechanism.METHODS:PANC-1 cells were cultured in vitro,and CCK8 method was used to detect the cell viability after different concentrations of JPHYD and GEM,and ap-propriate concentrations were selected for follow-up experiments.EDU assay and clonogenesis assay were used to detect cell proliferation.The apoptosis rate was detected by flow cytometry.Intracellular reactive oxygen species(ROS)were de-tected by DCFH-DA fluorescent probe and lipid peroxidation was detected by BODIPY 581/591C11 staining.The contents of glutathione(GSH),ferrous ion(Fe2+)and malondialdehyde(MDA)in the cells were detected by the kit.The mRNA levels and protein expression levels of Nrf2,HO-1,SLC7A11,GPX4,TFR1 and ACSL4 were detected by RT-qPCR and Western blot.RESULTS:Compared with the control group,the cell viability of PANC-1 treated with JPHYD and GEM was significantly decreased in a concentration-dependent manner.And the combined use of the two can significantly im-prove the cytotoxic effect of GEM and have a synergistic effect;Compared with control group,JPHYD group,GEM group and JPHYD+GEM group can significantly reduce EDU positive efficiency,colony formation numbers and promote cell apoptosis,and the combined group has the most obvious effect.After adding JPHYD+GEM into the cells,the cells be-came rounded and the cell viability decreased.The addition of ferrostatin-1(Fer-1),an inhibitor of ferroptosis,had no significant effect on cell morphology and viability,and the co-treatment with JPHYD+GEM and Fer-1 could reverse the ef-fects of JPHYD+GEM on cell morphology and viability.Compared with control group and GEM group,JPHYD+GEM group can significantly increase the levels of intracellular reactive oxygen species(ROS)and lipid peroxidation,increase the levels of Fe2+and MDA,decrease the levels of GSH,further promote lipid peroxidation and induce ferroptosis.JPHYD+GEM also significantly down-regulated the mRNA and protein expressions of Nrf2,HO-1,SLC7A11 and GPX4,and up-regulated the mRNA and protein expressions of TFR1 and ACSL4.The addition of Fer-1 significantly reversed the activation of iron death in the combined treatment group and reversed its efficacy,and the difference was statistically signif-icant.CONCLUSION:Jianpi Huayu decoction and gemcitabine may induce ferroptosis of PANC-1 cells by inhibiting Nrf2/SLC7A11/GPX4 axis in vitro,thus playing a synergistic anticancer role.
5.Accuracy of screw placement and learning curve analysis of robot-assisted minimally invasive transforaminal lumbar interbody fusion
Wenjie ZHONG ; Wenao LIAO ; Xilin LIU
Chinese Journal of Spine and Spinal Cord 2025;35(1):53-60
Objectives:To investigate the pedicle screw placement accuracy of robot-assisted minimally inva-sive transforaminal lumbar interbody fusion(RA-MIS-TLIF)and analyze its learning curve.Methods:A retro-spective analysis was conducted on the clinical data of 160 patients with degenerative lumbar diseases treated at the Department of Orthopedics,Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hos-pital between January 2019 and June 2022.The patients were divided into a robot group(n=80)and a manual group(n=80)based on the surgical approach.Each group was further divided into four subgroups(1,2,3,4)of 20 patients each,arranged sequentially according to the order of surgery dates.The total operative time,blood loss,and planning and screw placement time of the two surgical methods were statistically analyzed.Preoper-ative,postoperative 1d,and six-month postoperative visual analogue scale(VAS)scores for pain and Oswestry disability index(ODI)were compared across the four subgroups.The trends in total operative time and blood loss with increasing surgery cases were analyzed using a logarithmic regression model.Screw placement clas-sification was evaluated using the Gertzbein-Robbins classification based on postoperative CT scans,and then the screw placement accuracy was calculated.Results:All the surgeries were successfully completed.In the robot group,the total operative time was 162.9±5.7min,blood loss was 91.4±9.5mL,and planning and screw placement time was 42.1±1.3min;In the manual group,the total operative time was 169.1±6.6min,blood loss was 101.0±9.2mL,and planning and screw placement time was 57.0±6.3min.A total of 320 screws were placed in each group,with accuracy rates of 97%(311/320)in the robot group and 92%(295/320)in the manual group.Neither group had grade C or D screws.The total operative time decreased with the increase in the number of surgeries in both groups.The robot group achieved relative stability between subgroups 2 and 3[y=-5.894×ln(x)+183.891,R2=0.576,P<0.05],while the manual group achieved relative stability between subgroups 3 and 4[y=-4.424×ln(x)+184.221,R2=0.376,P<0.05].The blood loss also decreased with the increase in the number of surgeries in both groups,the robot group achieved relative stability between subgroups 2 and 3[y=-9.480×ln(x)+125.361,R2=0.547,P<0.05],and the manual group achieved relative stability also between subgroups 2 and 3[y=-3.868×ln(x)+114.183,R2=0.148,P<0.05].Postoperative VAS scores in the robot group decreased from 6.9±1.1 preoperatively to 2.4±0.9 on postoperative 1d(P<0.05)and to 1.1±0.4 at six months(P<0.05).In the manual group,VAS scores decreased from 7.0±0.9 preoperatively to 2.4±0.9 on postoperative 1d(P<0.05)and to 1.4±0.6 at six months(P<0.05).ODI in the robot group decreased from(59.5±7.1)%preoperatively to(20.0±4.1)%on postoperative 1d(P<0.05)and to(10.8±3.0)%at six months(P<0.05).In the manual group,ODI scores decreased from(57.7±6.9)%preoperatively to(19.6±4.6)%on postoperative 1d(P<0.05)and to(11.3±3.4)%at six months(P<0.05).All the 160 patients completed follow-ups with no severe complications reported during the follow-up period.Conclusions:Comparing with traditional MIS-TLIF with manual pedicle screw placement,RA-MIS-TLIF has a higher accuracy of screw placement and a smoother learning curve,which stabilizes after approximately 20 cases.
6.Accuracy of screw placement and learning curve analysis of robot-assisted minimally invasive transforaminal lumbar interbody fusion
Wenjie ZHONG ; Wenao LIAO ; Xilin LIU
Chinese Journal of Spine and Spinal Cord 2025;35(1):53-60
Objectives:To investigate the pedicle screw placement accuracy of robot-assisted minimally inva-sive transforaminal lumbar interbody fusion(RA-MIS-TLIF)and analyze its learning curve.Methods:A retro-spective analysis was conducted on the clinical data of 160 patients with degenerative lumbar diseases treated at the Department of Orthopedics,Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hos-pital between January 2019 and June 2022.The patients were divided into a robot group(n=80)and a manual group(n=80)based on the surgical approach.Each group was further divided into four subgroups(1,2,3,4)of 20 patients each,arranged sequentially according to the order of surgery dates.The total operative time,blood loss,and planning and screw placement time of the two surgical methods were statistically analyzed.Preoper-ative,postoperative 1d,and six-month postoperative visual analogue scale(VAS)scores for pain and Oswestry disability index(ODI)were compared across the four subgroups.The trends in total operative time and blood loss with increasing surgery cases were analyzed using a logarithmic regression model.Screw placement clas-sification was evaluated using the Gertzbein-Robbins classification based on postoperative CT scans,and then the screw placement accuracy was calculated.Results:All the surgeries were successfully completed.In the robot group,the total operative time was 162.9±5.7min,blood loss was 91.4±9.5mL,and planning and screw placement time was 42.1±1.3min;In the manual group,the total operative time was 169.1±6.6min,blood loss was 101.0±9.2mL,and planning and screw placement time was 57.0±6.3min.A total of 320 screws were placed in each group,with accuracy rates of 97%(311/320)in the robot group and 92%(295/320)in the manual group.Neither group had grade C or D screws.The total operative time decreased with the increase in the number of surgeries in both groups.The robot group achieved relative stability between subgroups 2 and 3[y=-5.894×ln(x)+183.891,R2=0.576,P<0.05],while the manual group achieved relative stability between subgroups 3 and 4[y=-4.424×ln(x)+184.221,R2=0.376,P<0.05].The blood loss also decreased with the increase in the number of surgeries in both groups,the robot group achieved relative stability between subgroups 2 and 3[y=-9.480×ln(x)+125.361,R2=0.547,P<0.05],and the manual group achieved relative stability also between subgroups 2 and 3[y=-3.868×ln(x)+114.183,R2=0.148,P<0.05].Postoperative VAS scores in the robot group decreased from 6.9±1.1 preoperatively to 2.4±0.9 on postoperative 1d(P<0.05)and to 1.1±0.4 at six months(P<0.05).In the manual group,VAS scores decreased from 7.0±0.9 preoperatively to 2.4±0.9 on postoperative 1d(P<0.05)and to 1.4±0.6 at six months(P<0.05).ODI in the robot group decreased from(59.5±7.1)%preoperatively to(20.0±4.1)%on postoperative 1d(P<0.05)and to(10.8±3.0)%at six months(P<0.05).In the manual group,ODI scores decreased from(57.7±6.9)%preoperatively to(19.6±4.6)%on postoperative 1d(P<0.05)and to(11.3±3.4)%at six months(P<0.05).All the 160 patients completed follow-ups with no severe complications reported during the follow-up period.Conclusions:Comparing with traditional MIS-TLIF with manual pedicle screw placement,RA-MIS-TLIF has a higher accuracy of screw placement and a smoother learning curve,which stabilizes after approximately 20 cases.
7.Jianpi-Huayu decotion combined with gemcitabine induces ferroptosis and inhibits growth of pancreatic cancer cells through Nrf2/SLC7A11/GPX4 axis
Xinqiu CHEN ; Zhengze ZHANG ; Shuwei LIU ; Xiaoyu ZHU ; Yongheng LAI ; Chongkai FANG ; Junhai HUANG ; Xilin ZHAO ; Chong ZHONG
Chinese Journal of Pathophysiology 2025;41(6):1077-1087
AIM:To investigate the effect of Jianpi-Huayu decoction(JPHYD)combined with gemcitabine(GEM)on ferroptosis of pancreatic cancer PANC-1 cells and its mechanism.METHODS:PANC-1 cells were cultured in vitro,and CCK8 method was used to detect the cell viability after different concentrations of JPHYD and GEM,and ap-propriate concentrations were selected for follow-up experiments.EDU assay and clonogenesis assay were used to detect cell proliferation.The apoptosis rate was detected by flow cytometry.Intracellular reactive oxygen species(ROS)were de-tected by DCFH-DA fluorescent probe and lipid peroxidation was detected by BODIPY 581/591C11 staining.The contents of glutathione(GSH),ferrous ion(Fe2+)and malondialdehyde(MDA)in the cells were detected by the kit.The mRNA levels and protein expression levels of Nrf2,HO-1,SLC7A11,GPX4,TFR1 and ACSL4 were detected by RT-qPCR and Western blot.RESULTS:Compared with the control group,the cell viability of PANC-1 treated with JPHYD and GEM was significantly decreased in a concentration-dependent manner.And the combined use of the two can significantly im-prove the cytotoxic effect of GEM and have a synergistic effect;Compared with control group,JPHYD group,GEM group and JPHYD+GEM group can significantly reduce EDU positive efficiency,colony formation numbers and promote cell apoptosis,and the combined group has the most obvious effect.After adding JPHYD+GEM into the cells,the cells be-came rounded and the cell viability decreased.The addition of ferrostatin-1(Fer-1),an inhibitor of ferroptosis,had no significant effect on cell morphology and viability,and the co-treatment with JPHYD+GEM and Fer-1 could reverse the ef-fects of JPHYD+GEM on cell morphology and viability.Compared with control group and GEM group,JPHYD+GEM group can significantly increase the levels of intracellular reactive oxygen species(ROS)and lipid peroxidation,increase the levels of Fe2+and MDA,decrease the levels of GSH,further promote lipid peroxidation and induce ferroptosis.JPHYD+GEM also significantly down-regulated the mRNA and protein expressions of Nrf2,HO-1,SLC7A11 and GPX4,and up-regulated the mRNA and protein expressions of TFR1 and ACSL4.The addition of Fer-1 significantly reversed the activation of iron death in the combined treatment group and reversed its efficacy,and the difference was statistically signif-icant.CONCLUSION:Jianpi Huayu decoction and gemcitabine may induce ferroptosis of PANC-1 cells by inhibiting Nrf2/SLC7A11/GPX4 axis in vitro,thus playing a synergistic anticancer role.
8.Allicin inhibits HBV replication through the HBV promoter SP2
Lili LU ; Xilin ZHU ; Xiaopan WU ; Ying LIU
Basic & Clinical Medicine 2025;45(4):465-470
Objective To investigate the effect of allicin on the replication of hepatitis B virus(HBV)and to pre-liminarily elucidate its underlying molecular mechanisms.Methods HepG 2.2.15 cells were treated with different concentrations of allicin and the levels of HBsAg and HBeAg were assessed by ELISA.Cell viability was evaluated using the CCK-8 assay to determine the optimal concentration of allicin;HepG2-NTCP cells were incubated with the optimal concentration of allicin for 48 hours,and the expression of HBV-related markers was detected by RT-qPCR;The activity of four HBV promoters(Enh I/Xp,SP1,SP2,and CP)was analyzed using a dual-lucif-erase reporter gene experiment.The effect of allicin on promoter activity was assessed;Gene-regulation tools were used to predict potential transcription factor that might bind to the promoter.After over-expressing the transcription factor,cells were incubated with allicin and the effect on promoter activity was examined;Finally,ChIP was used to confirm whether these transcription factors bind to the HBV promoters and whether allicin treatment affects this binding.Results Allicin significantly reduced the expression of HBsAg and slightly lowered the expression of HBeAg(P<0.001);A concentration of 40 μmol/L allicin significantly inhibited HBV DNA replication and tran-scription(P<0.05),without affecting cell viability;Allicin also significantly suppressed the activity of the HBV promoter SP2(P<0.001).Further investigation revealed that the transcription factor SP1 could bind to the DNA se-quence of the HBV promoter SP2,and this binding was significantly inhibited after allicin treatment(P<0.001).Conclusions Allicin inhibits the binding of the transcription factor SP1 to HBV promoter SP2,thereby reducing the transcriptional activity of HBV and suppressing viral replication.
9.Molecular mechanism of ATF6 regulating the reproduction related gene HSPA1L
Yuanyuan WANG ; Xilin ZHU ; Xiaopan WU ; Ying LIU
Basic & Clinical Medicine 2024;44(1):37-42
Objective To explore the effect of endoplasmic reticulum stress activating transcription factor 6(ATF6)on the expression of reproduction related gene heat shock protein family A member 1 like(HSPA1L)and preliminari-ly clarify its regulatory molecular mechanism.Methods The ATF6 over-expression plasmid was transfected into HEK-293T cells and the over-expression efficiency was verified by RT-qPCR and Western blot.The transcriptome sequen-cing information of testis tissue of male ATF6 knockout mice was used to screen five reproduction related genes down-stream of ATF6.The dual luciferase reporter assay selected the downstream genes with high promoter activity and de-tected the effect of over-expression of ATF6 on the promoter activity of downstream genes.The possible binding sites of ATF6 and downstream gene promoters were predicted by gene-regulation.RT-qPCR and Western blot were used to detect the effect of over-expression of ATF6 on downstream gene expression in HEK-293T cells.Whether ATF6 binds to downstream gene promoters was determined by electrophoretic mobility shift assay(EMSA).Results The expres-sion of ATF6 mRNA(P<0.001)and protein(P<0.001 and P<0.05)in HEK-293T cells was significantly increased after transfection.HSPA1L(P<0.001 and P<0.05),a reproductive related gene downstream of ATF6 was screened by transcriptome sequencing and dual luciferase reporter assay.ATF6 promoted the truncated promoter activity of HSPA1L(P<0.001).After over-expression of ATF6,the expression of HSPA1L was significantly increased(P<0.001).The differences were statistically significant.ATF6 protein could bind to the aagtcgtcac DNA sequence of HSPA1L promoter.Conclusions ATF6,a key molecule of endoplasmic reticulum stress,regulates the expression of reproduction related gene HSPA1L by binding to the promoter of HSPA1L.This result will lay a foundation for further research on the prevention or treatment of endoplasmic reticulum stress(ERS)related male infertility.
10.A qualitative study of effect of psychodrama therapy in adolescent inpatients with mental disorders
Yanru LIU ; Xilin WANG ; Huaqing LIU ; Xiaoming ZHANG ; Zhiren WANG
Chinese Mental Health Journal 2024;38(3):213-217
Objective:To explore the effect of psychodrama therapy in adolescent inpatients with mental disor-ders.Methods:Totally 22 patients with mental disorders(aged 12-18 years)were recruited from the inpatient children's ward of a certain hospital.According to the interview outline,a semi-structured interview was conducted in adolescent inpatients with mental disorders who participated in 4 sessions of psychodrama therapy.These patients took psychotropic drugs regularly.The interview data were analyzed in depth by using the grounded theory meth-od.The self-compiled Psychodrama Therapy Participation Feeling and Evaluation Questionnaire was used to investi-gate the feelings,gains or changes of patients after psychodrama therapy.Results:The interview analysis found that the effects of psychodrama therapy included reducing symptoms,improving self-esteem,promoting functional recov-ery and improving social adaptability.The results of the self-compiled Psychodrama Therapy Participation Feeling and Evaluation Questionnaire showed that the top 4 aspects were promoting interpersonal relationship,being inter-esting,willing to continue to participate and improving self-confidence.Conclusion:Psychodrama therapy has an auxiliary therapeutic effect in adolescent inpatients with mental disorders,and can promote the improvement of self-esteem and social skills.

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