1.Predictive value of contrast-enhanced ultrasound in evaluating delayed graft function in kidneys from donation after brain death
Jing SUN ; Yue WANG ; Jianlei JI ; Jinquan LIU ; Xiaodong WU ; Chuanshen XU ; Jianhong WANG
Organ Transplantation 2025;16(3):460-466
Objective To investigate the predictive value of quantitative parameters of contrast-enhanced ultrasound (CEUS) in evaluating kidneys from donation after brain death (DBD) for the occurrence of delayed graft function (DGF) in recipients. Methods The clinical data of 134 DBD donors and 202 corresponding kidneys and recipients were retrospective analyzed. The recipients were divided into DGF group (n=39) and non-DGF group (n=163) according to the renal function after kidney transplantation. Conventional ultrasound, CEUS parameters, and clinical data were compared between the two groups. Receiver operating characteristic (ROC) curves were used to determine the optimal cut-off values for predicting DGF using CEUS parameters, clinical parameters, and their combination, based on the highest Youden index. The predictive ability of different parameters for DGF was evaluated. Results There were statistically significant differences in cortical peak intensity (PIc), medullary peak intensity (PIm), donor albumin (ALB), serum creatinine (Scr) after admission, and the Na+ concentration of recipients between the two groups (all P<0.05). The area under the curve (AUC) for predicting DGF using the combination of CEUS parameters PIc and PIm was 0.711, with an optimal cut-off value of 0.193 and a Youden index of 0.382. The AUC for predicting DGF using the combination of CEUS parameters PIc, PIm and clinical parameters was 0.808, with an optimal cut-off value of 0.191 and a Youden index of 0.517. The sensitivity and specificity were 0.769 and 0.613 for the former, and 0.769 and 0.748 for the latter, respectively. The AUC for predicting DGF using CEUS parameters PIc and PIm combined with clinical parameters was significantly higher than that using CEUS parameters PIc and PIm (P<0.05). Conclusions The CEUS quantitative parameters PIc and PIm have good predictive value in assessing kidneys from DBD donors for DGF in recipients, and the diagnostic efficacy is better when combined with clinical parameters.
2.Immune reconstitution efficacy of human thymic slices transplanted to different parts of nude mice
Wanqing GUAN ; Guihua LUO ; Jingxuan HAN ; Jianhong MI ; Zeqing FENG ; Yuzhang WU
Immunological Journal 2024;40(2):113-121
This study was designed to explore the immune reconstitution efficacy of human thymic slices transplanted into renal capsule,subcutaneous or muscle of nude mice,and further explore the optimal location of heterotopic transplantation.The thymus tissue discarded from congenital heart disease patients was made into 0.5-1 mm thick tissue sections and cultured in vitro to remove immune cells.H&E staining and immunohistochemical staining were used to assess the residual tissue structure and cell types in thymic slices,while quantitative PCR methods were used to assess the function of residual cells in thymic slices.Then thymic slices were transplanted into the renal capsule,subcutaneous or muscle of nude mice,and the immune reconstitution efficacy was compared by flow cytometry and histology.Data showed that after 14 days of culture in vitro,the clearance rate of T lymphocytes in the thymic slices was more than 90%,and the epithelial cell network structure of the tissue was intact,while a large number of macrophages,dendritic cells and endothelial cells remained.Quantitative PCR results showed that the gene expression levels of epithelial cell markers and secreted cytokines in cultured thymic slices could be effectively maintained.Flow cytometry showed that at 16 weeks after transplantation,the proportion of T cells in peripheral blood of mice in different transplantation groups were significantly increased,whereas the proportion of T cells in muscle group was the highest.In situ histological examination showed that the regeneration of thymus tissue was detected at all three transplant sites.In addition,the graft detection rate was 40%in the renal capsule group,60%in the subcutaneous group and 100%in the musclegroup.In conclusion,the human thymic slices cultured in vitro for 14 days retain a complete thymic matrix microenvironment.Transplantation of human thymic slices can effectively reconstruct the ratio of T cells in nude mice,and the muscle is the most effective transplantation site.
3.Effect of Early Active Cycle of Breathing Technique Training on Aspiration in Patients with Dysphagia after Partial Laryngectomy
Wenliang XIE ; Jie TAN ; Jingang AI ; Jianhong WU ; Jing PENG
Journal of Audiology and Speech Pathology 2024;32(2):129-133
Objective To investigate the effect of early active cycle breathing technique(ACBT)on aspiration in patients with dysphagia after partial laryngectomy.Methods A total of 40 patients with laryngeal cancer with dysphagia who were hospitalized in the Department of Otorhinolaryngology of the Third Xiangya Hospital of Central South University in January 2019~January 2022 were selected,and the patients were randomly divided into 20 cases in the observation group and the control group by random number method,the control group was given routine swallowing function training,and the observation group was combined with active cycle of breathing technique(ACBT)on the basis of the control group.The two groups were treated 5 days a week,twice a day,45 minutes each for 2 weeks.The M.D.Anderson Dysphagia Inventory(MDADI),maximum phonation time(MPT),and Standardized Swallowing Assessment(SSA),flexible endoscopic examination of swallowing(FEES)combined with modified invasion and aspiration score(MPAS score)and overall clinical efficacy before and after treatment were compoued between the two groups.Results After 2 weeks of treatment,the swallowing function of both groups improved,but the MDADI scores in the observation group were better than those of in the control group in all cate-gories(P<0.001),MPT(7.19±1.31)was better than that of the control group(4.29±0.88)(=9.436,P<0.001),SSA(19.25±1.12)was better than that of the control group(21.20±2.55)(=-2.894,P<0.05),and FEES combined with MPAS score(1.75±0.85)was better than the control group(2.70±1.34)(=-2.674,P<0.001),and the overall clinical efficacy(18,90.00%)was better than the control group(12,60.00%)(Z=-3.894,P<0.001).Conclusion Early application of active breathing and circulation technique combined with swallowing training can improve the swallowing function of patients to a greater extent and reduce the incidence of aspiration compared with swallowing function training alone.
4.Research progress of lysine specific demethylase 1(LSD1)inhibitors
Xiaomeng ZHANG ; Jinyang FU ; Yanan HE ; Jianhong GONG ; Kun DU ; Ya WU ; Yanle ZHI
Journal of China Pharmaceutical University 2024;55(5):685-696
Lysine specific demethylase1(LSD1)is a flavin adenine dinucleotide(FAD)-dependent monoamine oxidase.Studies have confirmed that aberrant expression of LSD1 is closely related to tumor metastasis and proliferation,and is currently one of the important targets for tumor-targeted therapy.In addition,LSD1 is involved in the development of various conditions such as neurodegenerative diseases,cardiovascular diseases,and inflammatory responses.Currently,several inhibitors have been developed for the clinical research stage.In this paper,the structure and mechanism of action of LSD1 and the research progress of LSD1 inhibitors are briefly introduced to provide some reference for the design and development of novel LSD1 inhibitors.
5.Sulforaphane alleviates acute liver injury induced by diquat in mice by activating Keap1/Nrf2 signaling pathway
Jianhong WANG ; Liang PENG ; Liaozhang WU ; Shan HUANG ; Guoli HE ; Pei SHEN ; Jing LIANG ; Tingting HUANG ; Jiaming HUANG ; Hong ZHONG ; Manhong ZHOU
Chinese Critical Care Medicine 2024;36(11):1183-1189
Objective:To investigate the protective effect and possible mechanism of sulforaphane (SFN) on acute liver injury in mice induced by diquat (DQ) poisoning.Methods:Forty-eight male C57BL/6 mice were divided into Control group, DQ model group (DQ group), SFN intervention group (DQ+SFN group), and SFN control group (SFN group) using a random number table method, with 12 mice in each group. Acute liver injury mice model was established by one-time intraperitoneal injection of 1 mL of 40 mg/kg DQ solution at once. SFN group was injected with 1 mL of ddH 2O. After 4 hours of molding, 0.5 mL of 5 mg/kg SFN solution was injected into the peritoneal cavity of the DQ+SFN group and SFN group, once daily for 7 consecutive days. DQ group and Control group were injected with an equal amount of ddH 2O. Then, the mice were euthanized to collect liver tissue and blood samples, and the levels of plasma biomarkers alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as oxidative stress indicators such as superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) in liver tissue were measured. The changes of liver structure were observed under transmission electron microscopy. The apoptosis and reactive oxygen species (ROS) level in liver tissue were observed under fluorescence microscope. Western blotting was used to detect the protein expressions of nuclear factor E2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1), Kelch-like ECH-associated protein 1 (Keap1), and cleaved caspase-9 in liver tissue. Results:Compared with the Control group, the liver mitochondria in the DQ group showed severe swelling, partial dissolution of the matrix, and cristae rupture and loss; the levels of plasma AST and ALT significantly increased, the MDA content in the liver increased, the activities of SOD and GSH decreased, the level of ROS significantly increased, the number of apoptotic cells in the liver significantly increased, the protein expressions of Nrf2 and HO-1 significantly decreased, and the protein expressions of Keap1 and cleaved caspase-9 significantly increased. Compared with the DQ group, the mitochondrial damage in the DQ+SFN group was reduced, the levels of plasma AST and ALT were significantly reduced [ALT (U/L): 58.22±4.39 vs. 79.94±3.32, AST (U/L): 177.64±8.40 vs. 219.62±11.60, both P < 0.01], the liver MDA content decreased, and the activities of SOD and GSH increased [MDA (μmol/g: 5.63±0.18 vs. 5.96±0.29, SOD (kU/g): 102.05±4.01 vs. 84.34±5.34, GSH (mmol/g): 16.32±1.40 vs. 13.12±1.84, all P < 0.05], the production of ROS in liver tissue was significantly reduced [ROS (fluorescence intensity): 115.90±10.89 vs. 190.70±10.16, P < 0.05], and apoptotic cells were significantly reduced (cell apoptosis index: 4.39±1.00 vs. 10.71±0.56, P < 0.01), the protein expressions of Nrf2 and HO-1 were significantly increased, while the protein expressions of Keap1 and cleaved caspase-9 were significantly decreased (Nrf2/β-actin: 1.15±0.04 vs. 0.93±0.05, HO-1/β-actin: 1.75±0.12 vs. 0.78±0.04, Keap1/β-actin: 1.00±0.14 vs. 1.28±0.13, cleaved caspase-9/β-actin: 1.31±0.12 vs. 1.81±0.09, all P < 0.05). However, there was no statistically significant difference in various indicators between the SFN group and the Control group. Conclusion:SFN can activate the Keap1/Nrf2 signaling pathway to alleviate DQ induced acute liver injury in mice.
6.Analysis of antinuclear antibody in 9 528 pregnant women during early pregnancy in a hospital in Qingdao City
Shan KANG ; Guiliang HAO ; Xiangyu JING ; Mingzhen GUO ; Lin ZHAO ; Jianhong DONG ; Shuai WANG ; Menglu WU ; Mingran WU ; Qian XU ; Jinlian SONG
Chinese Journal of Preventive Medicine 2024;58(11):1727-1732
To analyze the positivity rate and titer of antinuclear antibody (ANA), as well as nuclear pattern and target antigen of ANA in healthy pregnant women during early pregnancy in Qingdao area. A prospective cohort study design was used to include a total of 9 528 healthy pregnant women registered at the Women and Children′s Hospital Affiliated to Qingdao University from March 2023 to June 2024.Indirect immunofluorescence assay (IIF) was used to detect ANA, its titer and cell staining pattern. Fifteen specific antibodies were tested using the magnetic bar code immunofluorescent luminescence method. Logistic regression model was used to analyze the risk factors of pregnancy with autoimmune disease(AID). The results showed that among 9 528 pregnant women in early pregnancy, 1 346 cases (14.1%) were positive of ANA, including 1 011 cases with a titer of 1∶100 (10.6%), 236 cases (2.5%) with a titer of 1∶320, and 99 cases (1.0%) were detected at a titer >1∶320. Among the 1 346 ANA-positive pregnant women, nuclear granular type accounted for the highest proportion (483 cases, 35.9%), followed by speckled type (347 cases, 25.8%) and cytoplasmic type (176 cases, 13.1%).Then, pregnant women with ANA titers ≥1∶100 were detected 15 specific antibodies.Anti-SSA was tested in 121 cases accounted for the majority, followed by 110 cases with anti-Ro-52, 56 cases with anti-SSB, 51 cases with anti-mitochondrial M2 subtype antibodies and 37 cases with anti-centromere B. In conclusion,in healthy pregnant women in Qingdao area, ANA positivity rate was 14.1%, and the titer of ANA was mainly at 1∶100.The predominant nuclear patterns were nuclear granular and speckled types.The specific autoantibodies were mainly anti-SSA antibodies and anti-Ro-52 antibodies.The detection of ANA and specific autoantibodies is of great significance for early prediction, diagnosis, and intervention of autoimmune diseases during pregnancy.
7.Biological properties of concentrated growth factor combined with bone marrow mesenchymal stem cell sheet and its effect on bone defect repairment
Jianhong SHI ; Yuanye TIAN ; Kai CHEN ; Gao SUN ; Guomin WU
Journal of Jilin University(Medicine Edition) 2024;50(6):1535-1546
Objective:To discuss the effect of concentrated growth factor(CGF)on the performance of bone marrow mesenchymal stem cells(BMSCs)sheets,and to clarify the role of CGF-containing composite cell sheets(CS)in the bone defect repairment.Methods:In in vitro experiments,the BMSCs were isolated and cultured from two 3-week-old SD rats;Alizarin Red S and Oil Red O staining were used to identify the osteogenic and adipogenic capabilities of BMSCs;CGF liquid extracts(CGFe)was prepared from three 3-week-old SD rats.The cells were divided into control group,traditional CS(BMSC-CS)group,and CGF-containing composite CS(CGF/BMSC-CS)group.The morphology of the CS in two groups was observed by HE staining.Alizarin Red and alkaline phosphatase(ALP)staining were used to detect the osteogenic differentiation of the CS in various groups;cell scratch assay was used to detect the migration abilities of the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the mRNA expression levels of ALP,collagen are type 1(COL-1),Runt-related transcription factor 2(RUNX2),and osteocalcin(OCN)in the cells in various groups.In in vivo experiments,15 SD rats were randomly divided into control group,BMSC-CS group,and CGF/BMSC-CS group;micro computed tomography(Micro-CT)was used to detect the bone formation parameters in skull defects of the rats in various groups;HE staining and Masson staining were used to observe the morphology of skull defect tissue of the rats in various groups.Results:The third-generation BMSCs were spindle-shaped,closely arranged,and grew in a vortex cluster.The Alizarin red staining results showed obvious calcium nodules,and the Oil red O staining showed red lipid droplets,confirming the cells'ability to undergo osteogenic and adipogenic differentiation.The CS were white and semi-transparent,with slightly curled edges.The peeled CS were irregularly curled and wrinkled.Compared with BMSC-CS group,the CS in CGF/BMSC-CS group were whiter,less transparent,significantly increased in thickness and extensibility,less prone to breakage,and had a certain degree of stickiness and plasticity.The HE staining results showed that compared with BMSC-CS group,the number of the cells of CS in CGF/BMSC-CS group was increased,with denser arrangement and more abundant extracellular matrix(ECM),which wrapped and connected the cells to form an integral sheet-like structure.The Alizarin red and ALP staining results showed that compared with control group,the ALP activity and mineralization uplift value of CS in BMSC-CS group were significantly increased(P<0.05);compared with control group and BMSC-CS group,the number of osteoblasts and red mineralized nodules in the CS in CGF/BMSC-CS group was significantly increased,with obvious deepening of the staining,increased positive area,and the ALP activity and mineralization uplift value were significantly increased(P<0.05).Compared with BMSC-CS group,the ALP activity and mineralization uplif value of the CS in CGF/DMSC-CS group were increased(P<0.05).The cell scratch assay results showed that after 24 of culture,compared with control group,the migration rates of the cells in BMSC-CS group and CGF/BMSC-CS group were significantly increased(P<0.05).Compared with BMSC-CS group,the migration rate of the cells in CGF/BMSC-CS group was significantly increased(P<0.01).After 48 h of culture,compared with control group,the migration rate of the cells in CGF/BMSC-CS group was significantly increased(P<0.05).The RT-qPCR results showed that compared with control group,the expression levels of COL-1 and OCN mRNA in the cells in BMSC-CS group were significantly increased(P<0.01),and the expression levels of ALP,COL-1,OCN,and RUNX2 mRNA in the cells in CGF/BMSC-CS group were significantly increased(P<0.01).Compared with BMSC-CS group,the expression levels of ALP,COL-1,and OCN mRNA in the cells in CGF/BMSC-CS group were significantly increased(P<0.01).The Micro-CT detection results showed that in control group,the boundary of the rat skull defect area was clear,with almost no new bone formation.In BMSC-CS group,a small amount of new bone formed only at the edge of the bone defect in skull of the rats,with a significant gap in the central area of the defect.In CGF/BMSC-CS group,new bone formed along the edge of the bone defect towards the central area in skull of the rats,repairing most of the bone defect.Compared with control group,the bone volume(BV)and trabecular number(Tb.N)of the rats in BMSC-CS group were significantly increased(P<0.05);the bone volume(BV),bone volume fraction[BV/tissue volume(TV)],trabecular thickness(Tb.Th),and trabecular number(Tb.N)in skull of the rats in CGF/BMSC-CS group,were significantly increased(P<0.05).Compared with BMSC-CS group,the BV,BV/TV,Tb.Th,and Tb.N in skull of the rats in CGF/BMSC-CS group were significantly increased(P<0.01).The HE and Masson staining observation showed that in control group,almost no new bone formed in the skull defect tissue of the rats,with only a large amount of collagen fibers connecting the two sides of the bone ends.In BMSC-CS group,a small amount of new bone formed only at the edge of the bone defect in skull tissue of the rats,with the central area of the defect containing dense collagen fibers connected to the newly formed bone at the defect edge.In CGF/BMSC-CS group,new bone tissue could be seen at the edge of the bone defect,and bone islands formed in the central area of the defect,surrounded by osteocytes and a large amount of collagen fibers.The Masson staining observation results showed that the cytoplasm and osteoid were red,and the collagen was blue.In CGF/BMSC-CS group,newly formed osteoid was observed in skull defect tissue of the rats,with the highest amount of new bone formation.Conclusion:CGF can promote the osteogenic differentiation and increase the richness of ECM in BMSCs sheets.CGF-containing composite CS can efficiently repair skull defects of the rats and serve as an ideal and safe material for promoting the bone regeneration.
8.Therapeutic effect of resveratrol on osteoarthritis of temporomandibular joint and its mechanism
Gao SUN ; Jing HE ; Qi ZHAO ; Jianhong SHI ; Zhiling LIAO ; Yuanye TIAN ; Guomin WU
Journal of Jilin University(Medicine Edition) 2024;50(6):1547-1556
Objective:To discuss the therapeutic effect of resveratrol on the temporomandibular joint osteoarthritis(TMJOA),and to clarify the related mechanism.Methods:Forty-five SD rats were randomly divided into control group,model group,and resveratrol group,and there were 15 rats in each group.The rats in model group and resveratrol group were intra-articularly injected with 50 μL of 20 g·L-1 monosodium iodoacetate(MIA)to set TMJOA rat models,while the rats in control group were injected with an equal volume of normal saline.Three weeks after modeling,the rats in resveratrol group received an injection of 80 μL resveratrol solution,once a week for three weeks,while the rats in control and model groups were injected with an equal volume of normal saline.Micro-computed tomography(Micro-CT)system was used to detect the condyle structure and the bone volume fraction(BV/TV),trabecular thickness(Tb.Th),trabecular spacing(Tb.Sp),and trabecular number(Tb.N)of the rats in various groups were calculated;HE staining and toluidine blue staining were used to observe the pathomorphology of temporomandibular joint(TMJ)tissue of the rats in various groups;immunohistochemistry was used to detect the expression levels of SRY-related HMG box(SOX)-9,matrix metalloproteinase(MMP)-13,silent information regulator(Sirt)1,phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),and phosphorylated mammalian target of rapamycin(p-mTOR)in TMJ tissue of the rats in various groups;real-time quantitative PCR(RT-qPCR)method was used to detect the expression levels of SOX-9,MMP-13,Sirt1,PI3K,mTOR,and Akt mRNA in TMJ tissue of the rats in various groups.Results:Three weeks after modeling,condylar bone was destructed,the surface was roughness,and continuity interruption were observed,indicating TMJOA model of the rats was established successfully.The Micro-CT system results showed that the condylar surface of the rats in control group was smooth and regularly shaped,with continuous bone texture;the rats in model group had significant condylar destruction,disrupted continuity,surface roughness,and varying degrees of bone defects;the rats in resveratrol group showed alleviated condylar lesions and improved appearance.Compared with control group,the BV/TV and Tb.Th of the rats in model group were significantly decreased(P<0.05),and Tb.Sp was significantly increased(P<0.05);compared with model group,the BV/TV and Tb.Th of the rats in resveratrol group were significantly increased(P<0.05),and the Tb.Sp was significantly decreased(P<0.05).The HE staining results showed clear layers and orderly chondrocyte arrangement in condyle of the rats in control group;the rats in model group showed rough uneven surface,obvious defects,and typical TMJOA features;the rats in resveratrol group showed slightly rough surface with generally clear layers and orderly arranged cells.The toluidine blue staining results showed distinct blue-purple staining of chondrocytes in hypertrophic layer of the rats in control group;pale staining or even loss of staining in some areas of the rats in model group;and distinct and relatively uniform staining in hypertrophic layer of the rats in resveratrol group.The immunohistochemistry results showed that compared with control group,the expression levels of MMP-13,PI3K,p-Akt,and p-mTOR proteins in TMJ tissue of the rats in model group were significantly increased(P<0.05),while the expression levels of SOX-9 and Sirt1 proteins in TMJ tissue of the rats were significantly decreased(P<0.05);compared with model group,the expression levels of SOX-9 and Sirt1 proteins in TMJ tissue of the rats in resveratrol group were significantly increased(P<0.05),whereas the expression levels of MMP-13,PI3K,p-Akt,and p-mTOR proteins were significantly decreased(P<0.05).The RT-qPCR results showed that compared with control group,the expression levels of MMP-13,PI3K,Akt,and mTOR mRNA in TMJ tissue of the rats in model group were significantly increased(P<0.05),while the expression levels of SOX-9 and Sirt1 mRNA were significantly decreased(P<0.05);compared with model group,the expression levels of SOX-9 and Sirt1 mRNA in TMJ tissue of the rats in resveratrol group were significantly increased(P<0.05),whereas the expression levels of MMP-13,PI3K,Akt,and mTOR mRNA were significantly decreased(P<0.05).Conclusion:Resveratrol has therapeutic effect on TMJOA,and its mechanism may be related to the activation of Sirt1 and inhibition of the PI3K-Akt-mTOR signaling pathway.
9.Single-center experience on 1 147 cases of liver transplantation
Xiaopeng XIONG ; Qingguo XU ; Xiaolong MIAO ; Hao WANG ; Peng LIU ; Xinqiang LI ; Xin ZHOU ; Xin WANG ; Feng WANG ; Yong ZHANG ; Yandong SUN ; Jinzhen CAI ; Liqun WU ; Chuanshen XU ; Jianhong WANG ; Qingjun GUO
Chinese Journal of General Surgery 2024;39(5):333-338
Objective:To analyze the prognostic outcomes of 1 147 patients who underwent liver transplantation at Qingdao University Affiliated Hospital and to summarize measures to enhance the efficacy of liver transplantation.Methods:A retrospective analysis was conducted on the clinical and follow-up data of 1 147 liver transplant patients at Qingdao University Affiliated Hospital.Results:The overall postoperative 1-, 3-, and 5-year survival rates for the 1 147 liver transplant patients were 87.20%, 73.40%, and 65.60%, respectively. The survival rates for benign disease liver transplant recipients were 88.01%, 84.98%, and 81.39% at 1, 3, and 5 years post-transplant, respectively, compared to recipients transplanted for malignancies of 78.11%, 64.41%, and 60.06% (all P<0.001). Among the mid vs more recent period, patients' 1-year and 3-year postoperative survival rates were 84.20%, 70.80% vs 90.50%, 71.70%, respectively,significantly in favor of recently enrolled patients ( P=0.022). In the complex surgery group, patients' 1-, 3-, and 5-year survival rates were 82.70%, 65.50%, 56.70%, while in less complicated group, it was 89.00%, 76.50%, 69.20% ( P<0.001). The primary causes of death for benign disease recipients were multi-organ failure (4.1%), while in recipients with malignant disease primary cause of death was tumor recurrence (23.7%). Postoperative complications included primary graft dysfunction, delayed graft function recovery, portal vein thrombosis, hepatic artery thrombosis, biliary stricture, post-transplant lymphoproliferative disorder, and graft-versus-host disease, with occurrence rates of 1.05%, 6.89%, 1.92%, 0.44%, 2.00%, 0.61%, and 0.44%, respectively. Conclusions:With the continuous improvement in surgical techniques and perioperative care levels, the 3-year survival rate of recipients at our center has increased. Malignant diseases and complex liver transplantation remain crucial factors affecting recipient prognosis, highlighting the need to further enhance comprehensive treatment capabilities for patients with malignant diseases and complex surgeries.
10.Effect of long non-coding RNA GHET1 on autophagy and drug resistance to cisplatin in head and neck squamous cell carcinoma
Shuping WU ; Jianhong YU ; Yu WU ; Hui LIU
Cancer Research and Clinic 2024;36(7):481-487
Objective:To investigate the effect of long non-coding RNA (lncRNA) GHET1 on autophagy and drug resistance to cisplatin in head and neck squamous cell carcinoma (HNSCC).Methods:Transcriptome sequencing (RNA-seq) data and the related clinical information of tumor samples from 504 HNSCC patients and 43 matched paracancerous tissues (> 2 cm from the tumor primary site margin) were downloaded from the Cancer Genome Atlas (TCGA) database. The data was updated in August 2022. R software was used to analyze the differences of GHET1 expression in cancer tissues, paracancerous tissues and 43 HNSCC matched samples. The data of 32 middle and advanced HNSCC patients who were eligible for surgery in Fujian Cancer Hospital from January 2019 to June 2023 were collected, and the differences of GHET1 expression between 14 patients insensitive to chemotherapy and 18 sensitive to chemotherapy were compared. Human HNSCC cell lines FaDu and Cal27 were selected to establish cisplatin-resistant HNSCC cell lines FADU-DDP-R and CAL27-DDP-R. The cells were transfected with siRNA targeting GHET1 (corresponding siRNA group), and the control group was transfected with negative control siNC. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of GHET1 and Beclin-1mRNA. The expressions of Beclin-1, LC3-Ⅰ/Ⅱ, p63 and GAPDH protein were detected by using Western blot. The autophagy inhibitor 3-methyladenosine (3-MA) was used to treat FADU-DDP-R and CAL27-DDP-R cell lines; the half inhibitory concentration ( IC50), the expression differences of GHET1, Beclin-1 mRNA and autophagy related protein of cisplatin were compared between the drug-resistant cell lines and the drug-resistant cell lines of the 3-MA group. Results:In TCGA database, the relative expression level of GHET1 in 504 HNSCC tissues was higher than that in 43 paracancerous tissues, and the difference was statistically significant ( Z = 2.57, P < 0.05); the relative expression level of GHET1 in 43 HNSCC tissues was higher than that in the paired paracancerous tissues, and the difference was statistically significant ( t = 3.24, P = 0.002). The relative expression level of GHET1 in HNSCC of 18 patients in chemotherapy-sensitive group and 14 patients in chemotherapy-insensitive group was 1.01±0.12 and 2.05±0.26, respectively, and the expression of GHET1 in the chemotherapy-insensitive group was higher than that in the chemotherapy-sensitive group ( t = 15.45, P < 0.001). IC50 of FaDu and FADU-DDP-R cell lines was (35.6±1.9) μmol/L and (86.2±2.7) μmol/L, respectively, and the difference was statistically significant ( t = 26.64, P < 0.001). The IC50 of Cal27 and CAL27-DDP-R cell lines was (68.8±8.9) μmol/L and (115.0±8.2) μmol/L, respectively, and the difference was statistically significant ( t = 6.60, P < 0.01). The relative expression levels of GHET1 in FaDu and FADU-DDP-R cell lines were 1.00±0.10 and 3.57±0.07, respectively ( t = 33.85, P < 0.001), and the relative expression level of GHET1 in FADU-DDP-R cell lines was higher than that in FaDu cell lines. The relative expression levels of GHET1 in Cal27 and Cal27-DDP-R cell lines were 1.00±0.08 and 2.06±0.11, respectively ( t = 13.25, P < 0.001), and the relative expression level of GHET1 in Cal27-DDP-R cell lines was higher than that in Cal27 cell lines. Western blot showed that the relative expression levels of autophagy related proteins Beclin-1, LC3-Ⅰ, LC3-Ⅱ and p63 in FADU-DDP-R and CAL27-DDP-R cell lines were higher than those in FaDu and Cal27 cell lines (all P < 0.05). The results of qRT-PCR showed that the relative expression levels of GHET1 in FADU-DDP-R cell lines in the control group and siGHET1 group were 1.00±0.12 and 0.20±0.06, respectively ( t = 10.52, P < 0.001). The relative expression levels of GHET1 in CAL27-DDP-R cell lines in the control group and siGHET1 group were 1.00±0.09 and 0.51±0.03, respectively ( t = 8.90, P < 0.001). The relative expression level of GHET1 in the knockdown group of the 2 cell lines was lower than that of the corresponding drug-resistant cell lines. The relative expression levels of Beclin-1 mRNA in FADU-DDP-R cell line in the control group and siGHET1 group were 1.00±0.09 and 0.60±0.07, respectively ( t = 6.08, P < 0.01); the relative expression levels of Beclin-1 mRNA in Cal27-DDP-R cell lines in the control group and siGHET1 group were 1.00±0.14 and 0.65±0.04, respectively ( t = 4.31, P < 0.05); the relative expression levels of Beclin-1 mRNA in drug-resistant cell lines after knocking down GHET1 were lower than those in corresponding drug-resistant cell lines. Western blot showed that the relative expression levels of Beclin-1, LC3-Ⅰ, LC3-Ⅱ and p63 in the knockdown group of drug-resistant cell lines were lower than those in the corresponding drug-resistant cells group. The cisplatin IC50 of drug-resistant cell lines in siGHET1 group was lower than that of the corresponding drug-resistant cell lines (all P < 0.05), and the cisplatin IC50 of drug-resistant cell lines in 3-MA group was lower than that of the corresponding drug-resistant cell lines (all P < 0.05). Conclusions:LncRNA GHET1 could induce the resistance to cisplatin by activating the autophagy in HNSCC.

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