1.AKT1-mediated autophagy of hepatocellular carcinoma cells enhances cell sensitivity to 125I seed irradiation
Chenyu WANG ; Zhizhou WU ; Li LIU ; Yunhua XIAO ; Xuequan HUANG
Journal of Army Medical University 2025;47(6):539-550
Objective To investigate the impact of serine/threonine-protein kinase 1(AKT1)-mediated autophagy in hepatocellular carcinoma(HCC)cells on their sensitivity to 125I seed irradiation.Methods ① iProX database and STRING12.0 website were utilized to analyze the proteomic data of HCC before and after 125I seed irradiation to explore the differentially expressed proteins and associated functional connections.Meanwhile,The Cancer Genomics Atlas(TCGA)database was employed to analyze the relationship between AKT1 expression level and the survival of HCC patients.② Human HCC cell lines HUH7 and Hep3B were exposed to continuous irradiation from 125I radioactive seeds with an initial apparent activity of 0.8 mCi per seed for approximately 120 h,accumulating a total dose of 8 Gy,while the control cells were cultured under normal condition for 120 h.③ Autophagy inhibitor,chloroquine(CQ)and inducer,rapamycin(RAPA)were used to treat the HCC cells respectively to establish the CQ group and the RAPA group.The lentiviral transfection technique was employed to construct the HCC cells with overexpressed AKT1,namely the AKT1 group.The HCC cells treated in the same way were continuously irradiated with 125I seeds for 120 h to construct the CQ+125I group,the RAPA+125I group,and the AKT1+125I group.④The changes in microtubule-associated protein light chain 3(LC3),p62,AKT1 and p-AKT1 were detected by Western blotting.Cell immunofluorescence assay was employed to observe the expression of autophagy related proteins,such as LC3.The colony forming ability and apoptotic rate were detected with plate cloning assay and flow cytometry.Results ① Continuous irradiation with 125I seeds resulted in decreased expression of p62 and increased ratio of LC3Ⅱ/LC3Ⅰ(P<0.05)when compared with the negative control(NC)group.Immunofluorescence assay revealed more green fluorescence spots of LC3.When compared with the 125I group,the CQ+125I group had significantly increased expression of p62(P<0.01),decreased ratio of LC3Ⅱ/LC3Ⅰ(P<0.01),lower apoptotic rate(P<0.01),and more colony formations(P<0.01).In contrast,the results in the RAPA+125I group were opposite to those of the CQ+125I group.② Analysis on the iProX database showed that the expression of AKT1 was decreased in the irradiated group,and analysis on the TCGA database indicated that high expression of AKT1 predicted a poor prognosis for HCC patients(P<0.01).③After irradiation with 125I seeds,the expression of AKT1 at both the RNA and protein levels was decreased in the 125I group(P<0.01).After overexpression of AKT1,the level of autophagy was decreased(P<0.05).Irradiation of HCC cells with overexpressed AKT1 using 125I seeds could partially restore the level of autophagy.In the AKT1+125I group,the expression of AKT1,pAKT1 and p62 were all decreased,and the ratio of LC3Ⅱ/LC3Ⅰwas increased than the AKT1 group(P<0.05).④ The apoptotic rate of the AKT1+125I group was lower than that of the 125I group(P<0.05)and higher than that in the AKT1 group(P<0.05).In HUH7 cells,the clonogenic ability of the AKT1+125I group was higher than that of the 125I group(P<0.05).In Hep3B cells,the clonogenic ability of the AKT1+125I group was higher than that of the 125I group,and the clonogenic ability of the AKT1 group was higher than that of the NC group(P<0.05).Conclusion 125I seed irradiation induce lethal autophagy in HCC cells by reducing the expression of AKT1,providing a new theoretical basis for the implantation of 125I radioactive seeds in the treatment of HCC.
2.Research progress on the application of human amniotic membrane in ocular fundus diseases
Fangxing ZHU ; Jiandong LIU ; Rongyu GAO ; Jiantao REN ; Xuequan SUN ; Xudong HUANG
Chinese Journal of Ocular Fundus Diseases 2025;41(4):324-328
Human amnion (hAM), as a biomaterial, has made significant progress in the field of ophthalmology, particularly in the treatment of retinal diseases. hAM possesses biological properties such as promoting tissue repair, inhibiting inflammation and neovascularization, and reducing fibrosis, which have led to its promising clinical outcomes in treating macular holes, retinal detachment, proliferative vitreoretinopathy, optic disc depression-related macular detachment, and age-related macular degeneration. The application of hAM can improve surgical success rates and promote vision recovery, with no significant rejection reactions observed due to its low immunogenicity. Nevertheless, the use of hAM still faces challenges in optimizing preparation and storage techniques, enhancing therapeutic efficacy, and reducing the risk of infectious disease transmission. Future research should focus on addressing these issues to further promote the application of hAM in retinal disease treatment and enhance its effectiveness.
3.Research progress on the application of human amniotic membrane in ocular fundus diseases
Fangxing ZHU ; Jiandong LIU ; Rongyu GAO ; Jiantao REN ; Xuequan SUN ; Xudong HUANG
Chinese Journal of Ocular Fundus Diseases 2025;41(4):324-328
Human amnion (hAM), as a biomaterial, has made significant progress in the field of ophthalmology, particularly in the treatment of retinal diseases. hAM possesses biological properties such as promoting tissue repair, inhibiting inflammation and neovascularization, and reducing fibrosis, which have led to its promising clinical outcomes in treating macular holes, retinal detachment, proliferative vitreoretinopathy, optic disc depression-related macular detachment, and age-related macular degeneration. The application of hAM can improve surgical success rates and promote vision recovery, with no significant rejection reactions observed due to its low immunogenicity. Nevertheless, the use of hAM still faces challenges in optimizing preparation and storage techniques, enhancing therapeutic efficacy, and reducing the risk of infectious disease transmission. Future research should focus on addressing these issues to further promote the application of hAM in retinal disease treatment and enhance its effectiveness.
4.Multi-slice spiral CT multi-planar reconstruction technology applicated in CT-guided puncture biopsy of pancreatic lesions
Liangyu DENG ; Xuequan HUANG ; Yunhua XIAO ; Chuang HE
Chinese Journal of Interventional Imaging and Therapy 2024;21(12):781-784
Objective To explore the application value of multi-slice spiral CT(MSCT)multi-planar reconstruction(MPR)technology for CT-guided puncture biopsy of pancreatic lesions.Methods A total of 104 patients with isolated pancreatic lesions who underwent CT-guided puncture biopsy were retrospectively enrolled,including 49 cases underwent MSCT-MPR assisted guided biopsy(MPR group)and 55 underwent MSCT-guided biopsy alone(non-MPR group).The success rate of puncture biopsy,the operating time,the times of needle adjustments,radiation dose-length product(DLP)and relative complications were recorded.Based on surgical pathology or follow-up(≥12 months)results,the sensitivity of biopsy was calculated.Results The success rate of one-time puncture was 100%in both groups.No significant difference of operating time was found between MPR([15.96±4.73]min)and non-MPR group([16.22±5.76]min,t=-0.250,P=0.803).MPR group had fewer times of needle adjustments([6.53±1.24]vs.[8.15±1.90],t=-5.201,P<0.001)and lower DLP([946.02±143.54]mGy·cm vs.[1 095.53±338.50]mGy·cm,t=-2.988,P=0.004)than non-MPR group.During biopsy,minor bleeding occurred in 1 case of MPR group and 5 of non-MPR group,then all spontaneously improved without special treatment.The diagnostic sensitivity of biopsy pathology was 95.56%(43/45)in MPR group and 92.16%(47/51)in non-MPR group.Conclusion MSCT-MPR technology could improve the accuracy of CT-guided puncture biopsy of pancreatic lesions and reduce radiation dose.
5.Application of automatic laser navigation in percutaneous bone marrow puncture biopsy
Junru XIONG ; Chuang HE ; Xuequan HUANG
Chongqing Medicine 2024;53(11):1636-1639
Objective To investigate the advantages and disadvantages of percutaneous bone marrow puncture biopsy guided by automatic laser navigation and free-hand percutaneous bone marrow biopsy guided by CT.Methods Seventy patients with bone marrow biopsy at the department of nuclear medicine in this hos-pital from January to June 2022 were selected as the study subjects and divided into the laser navigation group (bone marrow biopsy guided by automatic laser navigation,n=30) and free-hand puncture group (percutane-ous bone marrow biopsy guided by CT,n=40) based on different guidance methods.The duration of the pro-cedure,total radiation dose,total number of scans,positive rate of bone lesion biopsy and puncture complica-tions occurrence were compared between the two groups.Results Compared to the free-hand puncture group,the operation duration in the laser navigation group was shorter[11.00(7.00,13.00)min vs. 13.00(11.00,17.75)min],the total radiation dose was lower[439.00(291.00,568.75)mGy vs. 510.00(431.25,708.50) mGy],total number of scans was less[4.50(3.00,6.00)times vs. 6.00(5.00,7.00)times],and the differ-ences were statistically significant (P<0.05).There was no statistically significant difference in the positive rate of bone lesion biopsy between the laser navigation group and free-hand puncture group[96.67%(29/30) vs. 85.00%(34/40),P>0.05].No puncture complications occurred in both groups.Conclusion Automatic laser navigation could assist the operators to better complete percutaneous bone marrow biopsy procedures.
6.PGRMC1-mediated autophagy decreases the sensitivity of hepatocellular carcinoma cells to 125I particle irradiation
Pingping LIU ; Chenyu WANG ; Yunhua XIAO ; Chuang HE ; Junru XIONG ; Liangyu DENG ; Xuequan HUANG
Journal of Army Medical University 2024;46(9):1015-1023
Objective To investigate the effect of progesterone receptor membrane component 1(PGRMC1)mediated autophagy on the sensitivity of liver cancer cells to 125I particles irradiation.Methods Hepatoma cell lines Huh7 and LM3 were exposed to different doses(0,2,4,6 and 8 Gy)of 125I particles,and cell autophagy was observed by transmission electron microscopy(TEM).Then,autophagy inhibitor chloroquine(CQ),agonist rapamycin(Rapa),and PGRMC1 inhibitor AG-205 were used respectively to verify that PGRMC1-mediated autophagy plays a key role in the sensitivity of hepatocellular carcinoma cells to 125I particle irradiation.Cell proliferation,colony formation and apoptosis were detected by CCK-8 assay,clonal formation test and flow cytometry,respectively.The expression levels of PGRMC1,microtubule-associated protein light chain 3-Ⅰ(LC3-Ⅰ),LC3-Ⅱ and p62 were detected by Western blotting.Results Different doses of 125I particles irradiation significantly decreased the proliferation and clonogenesis of Huh7 and LM3 cells(P<0.05),and increased the apoptotic cells(P<0.01),in a dose-dependent manner.Compared with the 0 Gy group,the ratio of LC3-Ⅱ/LC3-Ⅰ in Huh7 and LM3 cells was obviously increased,and the expression of p62 was significantly down-regulated in the 6 Gy group.The proliferation capacity and clonal formation ability of Huh7 and LM3 cells were decreased significantly,and their apoptotic cells were increased notably in the 6 Gy+CQ group than the 6 Gy group,while the above results were on the contrary in the 6 Gy+Rapa group.The 6 Gy+AG205 group had notably decreased LC3-Ⅱ/LC3-Ⅰ ratio in the Huh7 and LM3 cells,up-regulated p62 expression,reduced cell proliferation capacity and clone formation ability,and enhanced cell apoptosis when compared with the 6 Gy group,and the above results of the 6 Gy+PGRMC1 group were opposite.Conclusion Increment of PGRMC1 induced by 125I irradiation can promote autophagy,increase the proliferation and clonogenesis,and reduce the apoptosis in hepatocellular carcinoma cells.
7.Multi-slice spiral CT multi-planar reconstruction technology applicated in CT-guided puncture biopsy of pancreatic lesions
Liangyu DENG ; Xuequan HUANG ; Yunhua XIAO ; Chuang HE
Chinese Journal of Interventional Imaging and Therapy 2024;21(12):781-784
Objective To explore the application value of multi-slice spiral CT(MSCT)multi-planar reconstruction(MPR)technology for CT-guided puncture biopsy of pancreatic lesions.Methods A total of 104 patients with isolated pancreatic lesions who underwent CT-guided puncture biopsy were retrospectively enrolled,including 49 cases underwent MSCT-MPR assisted guided biopsy(MPR group)and 55 underwent MSCT-guided biopsy alone(non-MPR group).The success rate of puncture biopsy,the operating time,the times of needle adjustments,radiation dose-length product(DLP)and relative complications were recorded.Based on surgical pathology or follow-up(≥12 months)results,the sensitivity of biopsy was calculated.Results The success rate of one-time puncture was 100%in both groups.No significant difference of operating time was found between MPR([15.96±4.73]min)and non-MPR group([16.22±5.76]min,t=-0.250,P=0.803).MPR group had fewer times of needle adjustments([6.53±1.24]vs.[8.15±1.90],t=-5.201,P<0.001)and lower DLP([946.02±143.54]mGy·cm vs.[1 095.53±338.50]mGy·cm,t=-2.988,P=0.004)than non-MPR group.During biopsy,minor bleeding occurred in 1 case of MPR group and 5 of non-MPR group,then all spontaneously improved without special treatment.The diagnostic sensitivity of biopsy pathology was 95.56%(43/45)in MPR group and 92.16%(47/51)in non-MPR group.Conclusion MSCT-MPR technology could improve the accuracy of CT-guided puncture biopsy of pancreatic lesions and reduce radiation dose.
8.Efficacy and prognostic factors analysis of CT-guided 125I seeds implantation for primary hepatocellular carcinoma
Qianqian YUAN ; Miaomiao HU ; Yanli MA ; Yuqing SONG ; Chuang HE ; Xuequan HUANG ; Chongshuang YANG ; He ZHU ; Zhe WANG ; Kaixian ZHANG ; Junjie WANG ; Jiuyan ZHANG ; Bin LIU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2022;42(11):666-671
Objective:To evaluate the efficacy and prognostic factors of 125I seeds implantation for primary hepatocellular carcinoma. Methods:From December 2011 to January 2021, 102 primary hepatocellular carcinoma patients (86 males, 16 females; median age 61 years) who underwent 125I seeds implantation from 5 hospitals in China were enrolled in this retrospective study. Local progression-free survival (LPFS), overall survival (OS) and the prognostic factors were analyzed. Kaplan-Meier method was used to draw the distribution curve of survival time, and LPFS rate and OS rate were calculated. Log-rank test and Cox regression were used to analyze the influencing factors of survival. Results:The median follow-up time was 38 months until April 2021. The local control rate was 96.1%(98/102). The 1-, 3- and 5-year LPFS rate were 61.3%, 25.5% and 12.7%, and the 1-, 3- and 5-year OS rate were 73.9%, 39.1% and 22.6%, respectively. There were 75 patients with progressive disease, including 42 patients with intrahepatic recurrence and metastasis after seed implantation, and 55 patients died. Multivariate analyses showed that short-term efficacy complete response (CR) (hazard ratio ( HR)=0.34, 95% CI: 0.20-0.58) was protective factor related to LPFS; short-term efficacy CR ( HR=0.25, 95% CI: 0.13-0.47) was the protective factors related to OS; Barcelona clinic liver cancer (BCLC) C stage ( HR=2.33, 95% CI: 1.27-4.27), intrahepatic progression and extrahepatic metastasis ( HR=3.18, 95% CI: 1.28-7.86; HR=3.23, 95% CI: 1.27-8.21) were independent risk factors related to OS. No sever adverse effects were observed. Conclusions:125I seeds implantation is safe and effective for the treatment of primary hepatocellular carcinoma. BCLC stage, short-term efficacy and post-implantation progression are independent factors related to survival time.
9.Experimental study of 125I seeds inhibiting angiogenesis of transplanted hepatocellular carcinoma
Chongshuang YANG ; Pingping LIU ; Xuequan HUANG ; Yunhua XIAO ; Henan LIU ; Liangshan LI ; Tingyuan LI
Chinese Journal of Radiological Medicine and Protection 2021;41(11):808-812
Objective:To investigate the effect of radioactive 125I seed on angiogenesis of subcutaneously transplanted hepatocellular carcinoma in nude mice and underlying mechanism. Methods:The subcutaneous transplanted tumor model of human hepatocellular carcinoma Huh7 cells was established in nude mice. Twelve nude mice were randomly divided into observation group and control group with 6 mice in each group. The 125I seed with activity of 2.96×10 7Bq was implanted into the transplanted tumor of observation group and another with 0 Bq as control group, respectively. The volume of the transplanted tumor was measured every 4 d and the growth curve of the tumor was recorded. The microvessel density (MVD) of the transplanted tumor was evaluated by immunohistochemical detection of CD31. VEGF-A and HIF-1α protein and mRNA were detected by immunohistochemistry and RT-PCR, respectively. Results:The growth rate of tumor in the observation group was slower than that in the control group, and the difference of tumor volume between two groups at 12 d after 125I seed implantation was significantly different( t=3.167, P<0.05). At 28 d after transplantation, the tumor volumes of control and observation group approached to (963.61 ± 89.56) mm 3and (602.10±75.98) mm 3, respectively. The MVD of the observation group was significantly lower than that of the control group ( t=6.361, P<0.05). The relative expression of VEGF-A and HIF-1α mRNA in the observation group was significantly lower than that in the control group ( t=10.480, 6.414, P<0.05). Protein expression levels of VEGF-A and HIF-1α in the observation group were lower than those in the control group ( t=10.890, 12.250, P<0.05). Conclusions:Radioactive 125I seed can inhibit the growth of HCC xenografts by reducing tumor microvessels, which may be related to the decrease of VEGF-A and HIF-1α expression.
10.Emergency establishment and application of biosafety autopsy and pathology platform
Pengnan ZHAO ; Xiaohong YAO ; Zongxing ZHANG ; Jiancheng QI ; Xuequan HUANG ; Zhicheng HE ; Ping CHEN ; Li HAN ; Dixiong XU ; Sibing ZHANG ; Xiuwu BIAN ; Hongyan ZHANG
Chinese Journal of Medical Science Research Management 2021;34(2):100-105
Objective:Through the establishment and application of the biosafety autopsy pathology platform in Huoshenshan Hospital, the feasibility and application effect of the biosafety autopsy pathology platform were analyzed.Methods:The feasibility and application effect of the biosafety autopsy pathology platform were analyzed by layout design, instruments and equipment preparation, testing methods examination, and effect evaluation.Results:A total number of 26 cases of systematic autopsy and 8 cases of minimally invasive autopsy (puncture) were performed on the biosafety autopsy pathology platform, and no one was infected. Some pathology original findings were identified, including COVID-19 and pathological characteristics of identification, SARS respiratory failure mechanism and treatment significance, systemic distribution and spreading mechanism of the new coronavirus, the " storm" of inflammation pathological basis, some tumor markers rise in pulmonary pathological cell source and the overcast with fibrosis characteristics, such findings play important roles in the clinical diagnosis and treatment of COVID-19.Conclusions:The study of autopsy pathology is of great significance for the prevention and control of emerging infectious disease, which calls for early intervention. To promote the standard construction of biosafety autopsy platform is the key to the pathological study of emerging infectious diseases. Pathological research and clinical diagnosis and treatment should be combined to inform each other.

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