1.Disulfidptosis-related lncRNAs establish new prognostic features and predict immunotherapeutic response in pancreatic cancer
Mingzheng TANG ; Xiaofeng LI ; Yao RONG ; Zhihang WU ; Guorong MA ; Songhua LIU ; Hui CAI
Journal of China Medical University 2024;53(1):20-26
Objective To screen long non-coding RNA(lncRNA)associated with disulfidptosis and investigate the immune landscape between lncRNA and pancreatic cancer,for effective guidance in clinical practice.Methods The normal and pancreatic cancer tissue samples were obtained from The Cancer Genome Atlas database,and the lncRNA associated with disulfidptosis was identified based on the Cox and LASSO regression analyses.A risk prognosis model was constructed,and its predictive performance was verified using comprehensive methods.An accurate nomogram was construted to predict the prognosis of patients with pancreatic cancer.The biological differences were analyzed via Gene Ontology,Gene Set Enrichment Analysis,and an immunoassay.The immunotherapy response was estimated using the tumor mutational burden(TMB)score.Results A total of 251 disulfidptosis-related lncRNAs were successfully identified,and three groups of lncRNAs were selected as the reference for the risk model.Pathway analysis showed that immune-related pathways were associated with disulfidptosis-related lncRNA risk models.The risk score was significantly correlated with immune cell infiltration and the ESTIMATE score.Patients with higher risk scores had elevated TMB,indicating that high-risk patients exhibited a better immune checkpoint blockade response.Conclusion The findings of this study contribute to a deeper understanding of disulfidpto-sis-related lncRNA and provide a potential therapeutic strategy for pancreatic cancer.
2.Application of Digital Galvanometer Scanner System for CO2 Fractional Laser Safety Improvement
Ming LI ; Wei FENG ; Songhua LIU
Chinese Journal of Medical Instrumentation 2024;48(3):323-329
An analog PID controller-based galvanometer scanner is widely used by fractional laser medical equipment(FLME)to scan lasers across tissue surfaces,achieving the desired therapeutic effect.This type of driver,primarily composed of passive components and operational amplifiers,can only accept commands from the central controller of the FLME,with a simple hardware circuit-based fault diagnosis;thus,the safety of the FLME is compromised.To address these issues,the failure mechanisms of galvanometers and their impact on the safety of FLME are thoroughly analyzed first.Then,an adaptive limit protection method,a coil open circuit fault diagnosis,a communication timeout protection based on two handshakes,and a galvanometer control timeout protection are proposed,respectively,based on a digital driver platform,to supplement the deficiencies in the original fault diagnosis and protection system.This ensures the safety of the FLME.Finally,the effectiveness of the proposed strategies is validated through experiments.
3.FBXO43 expression in pan-cancer and its relationship with immune cell infiltration and prognosis
Songhua LIU ; Yao RONG ; Mingzheng TANG ; Yunyang CHEN ; Pengfei LI ; Xiaojun YANG
Journal of China Medical University 2024;53(9):834-844
Objective To investigate the expression of FBXO43in various cancers and its relationship with immune cell infiltration and prognosis using bioinformatics.Methods Gene expression data for 33 cancers were obtained from The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases to analyze FBXO43expression.Clinical and survival data were sourced from the TCGA database.Gene Expression Profiling Interactive Analysis(GEPIA)was used to assess the correlation between FBXO43expression and the clinical stage.Kaplan-Meier survival analysis was used to evaluate the relationship between FBXO43expression and prognosis.R lan-guage was used to analyze the associations between FBXO43expression and clinical stages,immune cell infiltration,immune checkpoint genes,tumor mutation burden(TMB),microsatellite instability(MSI),and mismatch repair(MMR)genes.The potential biological mech-anisms of FBXO43were explored using gene set enrichment analysis(GSEA).Moreover,qRT-PCR was performed to measure FBXO43 expression in normal gastric mucosal cells(GES-1),gastric cancer cells(HGC-27,MGC-803,and MKN-45),normal liver cells(LO-2),and liver cancer cells(SMMC-7721,HEPG2,HuH7,and MHCC97-H).Results Combined TCGA and GTEx database statistics revealed higher FBXO43expression in 26 types of cancer tissues,including adrenocortical carcinoma(ACC),bladder urothelial carcinoma,breast invasive carcinoma,cervical squamous cell carcinoma and endocervical adenocarcinoma,cholangiocarcinoma,colon adenocarcinoma,and diffuse large B-cell lymphoma,than in normal tissues(P<0.05).However,FBXO43expression was lower in three types of cancer tissues:kidney chromophobe(KICH),testicular germ cell tumor(TGCT),and thyroid carcinoma(P<0.05).GEPIA data analysis showed that FBXO43expression positively correlated with the clinical stages of ACC,KICH,kidney renal clear cell carcinoma(KIRC),and kidney renal papillary cell carcinoma(KIRP)and negatively correlated with the clinical stages of ovarian serous cystadenocarcinoma(OV)and TGCT(P<0.05).Kaplan-Meier survival analysis indicated that abnormal FBXO43expression was associated with the prognosis of various cancers(P<0.05).Specifically,high FBXO43expression was a risk factor for ACC,KICH,KIRC,KIRP,low-grade glioma,liver hepato-cellular carcinoma,mesothelioma,and sarcoma,but protective in thymoma(THYM).The XCELL algorithm found that FBXO43expres-sion was negatively correlated with immune scores in nine types of cancer tissues,including glioblastoma multiforme,KIRP,acute myeloid leukemia,lung squamous cell carcinoma,and OV,and closely related to immune cell infiltration levels in 24 types of cancer tissues,espe-cially showing a significant negative correlation with most immune cells in SARC(P<0.01).Correlation analysis revealed a significant association between FBXO43and TMB,MSI,and MMR in all cancers.GSEA analysis indicated that FBXO43is involved in the cell cycle and immune-related functions in various tumors.qRT-PCR results showed that FBXO43expression was upregulated in liver cancer cells and downregulated in gastric cancer cells(all P<0.05).Conclusion Abnormal FBXO43expression is closely associated with the occur-rence and progression of multiple cancers.Thus,FBXO43may serve as a novel marker of immune cell infiltration and prognosis,thereby offering new directions for targeted cancer therapy.
4.Value of Deep Learning Ultrasound Radiomics Nomogram to Assess Invasive Metastasis in Invasive Breast Cancer
Songhua LI ; Chaojun WU ; Dayou WEI ; Shaofeng LI ; Youshi LUO ; Yan LIN ; Linyong WU
Chinese Journal of Medical Imaging 2024;32(8):803-808
Purpose To explore the value of deep learning ultrasound radiomics nomogram in assessing the biological characteristics of invasive metastases in invasive breast cancer.Materials and Methods A retrospective collection of ultrasound imaging data from 180 pathologically confirmed invasive breast cancer between January 2021 to December 2022 in Maoming People's Hospital was conducted,with pathological reports indicating the status of lymph node metastasis(LNM),lymphovascular space invasion(LVSI)or perineural invasion(PNI),according to the LNM/LVSI/PNI status,the three indexes were divided into the training cohort and the verification cohort by 8∶2.Based on Pyradiomics and ResNet50 deep learning extractor,1 316 radiomic features and 2 048 deep learning features were extracted,respectively.The random forest machine learning algorithm was employed to develop evaluation models,and the model scores were calculated.The deep learning radiomics nomograms were developed based on the radiomic and deep learning model scores.The receiver operating characteristic curve was used to assess the performance of the models.The Delong test was applied to analyze the performance differences between different models.Results In the evaluation of LNM,LVSI and PNI status,the area under the curve of all the nomogram in the cohorts demonstrated moderate or above assessment performance(≥0.73),with accuracies all greater than 0.70.Specifically,in the LNM evaluation,the area under the curve of the training cohort was 0.97,the accuracy was 0.93,the sensitivity was 0.88 and the specificity was 0.96.Through the Delong test,the assessment performance of the nomograms was superior to the radiomics models(LNM,Z=2.04,P=0.04;LVSI,Z=2.80,P=0.01;PNI,Z=3.52,P<0.01),and was superior to or similar to the deep learning models(LNM,Z=4.52,P<0.01;LVSI,Z=1.86,P=0.06;PNI,Z=0.31,P=0.76)in the training cohort.Conclusion The deep learning radiomics nomogram can effectively evaluate the biological characteristics of invasion and metastasis in invasive breast cancer.The nomogram improves the assessment performance by integrating the radiomic and deep learning feature information.
5.Marsdenia tenacissima injection induces the apoptosis of prostate cancer by regulating the AKT/GSK3β/STAT3 signaling axis.
Xiaolan LI ; Songhua HE ; Wei LIANG ; Weiquan ZHANG ; Xin CHEN ; Qiaofeng LI ; Xin YANG ; Yanying LIU ; Dan ZHU ; Li LI ; Buming LIU ; Zhiheng SU ; Jie CHEN ; Hongwei GUO
Chinese Journal of Natural Medicines (English Ed.) 2023;21(2):113-126
Marsdenia tenacissima injection, a standard Marsdenia tenacissima extract (MTE), has been approved as an adjuvant therapeutic agent for various cancers. Our previous study showed that MTE inhibited the proliferation and metastasis of prostate cancer (PCa) cells. However, the underlying mechanisms and active ingredients of MTE against PCa were not completely understood. This study revealed that MTE induced significant decreases in cell viability and clonal growth in PCa cells. In addition, MTE induced the apoptosis of DU145 cells by reducing the mitochondrial membrane potential and increasing the expression of Cleaved Caspase 3/7, Cyt c, and Bax. In vivo, DU145 xenografted NOD-SCID mice treated with MTE showed significantly decreased tumor size. TUNEL staining and Western blot confirmed the pro-apoptotic effects of MTE. Network pharmacology analysis collected 196 ingredients of MTE linked to 655 potential targets, and 709 PCa-associated targets were retrieved, from which 149 overlapped targets were screened out. Pathway enrichment analysis showed that the HIF-1, PI3K-AKT, and ErbB signaling pathways were closely related to tumor apoptosis. Western blot results confirmed that MTE increased the expression of p-AKTSer473 and p-GSK3βSer9, and decreased the expression of p-STAT3Tyr705in vitro and in vivo. A total of 13 compounds in MTE were identified by HPLC-CAD-QTOF-MS/MS and UPLC-QTOF-MS/MS. Molecular docking analysis indicated that six compounds may interact with AKT, GSK3β, and STAT3. In conclusion, MTE induces the endogenous mitochondrial apoptosis of PCa by regulating the AKT/GSK3β/STAT3 signaling axis, resulting in inhibition of PCa growth in vitro and in vivo.
Mice
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Animals
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Male
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Humans
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Mice, Inbred NOD
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Mice, SCID
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Marsdenia
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Proto-Oncogene Proteins c-akt
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Glycogen Synthase Kinase 3 beta
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Molecular Docking Simulation
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Phosphatidylinositol 3-Kinases
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Tandem Mass Spectrometry
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Prostatic Neoplasms
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Apoptosis
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STAT3 Transcription Factor
6.Effect of ultrasound-guided modified anterior approach block of subcostal lumbar quadratus muscle on postoperative analgesia in gynecological tumor patients
Ling LI ; Songhua LIU ; Yuan HUANG ; Jiahui ZHAO ; Lu LI ; Zhigang CHENG
Journal of Chinese Physician 2023;25(11):1619-1623
Objective:To evaluate the effect of ultrasound guided modified-subcostal approach to anterior quadratus lumborum block (MSC-AQLB) on postoperative pain relief in open gynecological tumor surgery.Methods:Fifty patients with open gynecological tumor surgery admitted to the Changsha Central Hospital from June 2022 to March 2023 were selected, aged 35-70 years old, with American Society of Anesthesiologist (ASA) grades Ⅰ-Ⅱ. They were randomly divided into two groups using a random number table: an improved subcostal lumbar quadratus anterior block combined with general anesthesia group (MQ group) and a simple general anesthesia group (GA group), with 25 patients in each group. Before induction of general anesthesia, the MQ group received bilateral MSC-AQLB under ultrasound guidance, with 20 ml of 0.4% ropivacaine administered to both sides; The GA group did not receive nerve block. Both groups received intravenous inhalation combined with general anesthesia during the surgery, and both groups received patient-controlled intravenous analgesia (PCIA) with sufentanil after the surgery. The block plane of the MQ group at 5 and 15 minutes after block was recorded, as well as the resting and active (cough) Visual Analogue Scale (VAS) of patients in both groups at extubation (T 1), departure from post anesthesia care unit (PACU) (T 2), postoperative 6 hours (T 3), 12 hours (T 4), 24 hours (T 5), and 48 hours (T 6), the effective and total compressions of the analgesic pump within 48 hours after surgery, the analgesic recovery rate, and postoperative patient satisfaction, the incidence of nausea and vomiting, complications related to nerve block (local anesthetic poisoning, muscle weakness, pneumothorax, bleeding, accidental entry into the abdominal cavity, kidney damage, etc.)were also recorded. Resultsl:The highest and lowest blocking planes of bilateral MSC-AQLB under ultrasound guidance were T 6 and L 1 (at 5 minutes), T 5 and L 2 (at 15 minutes), respectively. The resting and active VAS scores of the MQ group at T 1 to T 6 were significantly lower than those of the GA group (all P<0.05), and the effective press frequency, total press frequency, and analgesic recovery rate of the analgesic pump within 48 hours were significantly lower than those of the GA group (all P<0.05). The postoperative analgesic satisfaction score was higher than that of the GA group ( P<0.05), and the incidence of nausea and vomiting within 48 hours after surgery was significantly lower than that of the GA group ( P<0.05). The MQ group of patients did not experience complications related to nerve block such as kidney injury and muscle weakness. Conclusions:In open gynecological tumor surgery, ultrasound guided MSC-AQLB can effectively reduce postoperative pain scores, reduce the dosage of postoperative analgesics, reduce the incidence of nausea and vomiting, and significantly improve patient satisfaction with pain relief.
7.Effect of ultrasound-guided lumbar quadratus muscle block on the analgesic effect during and after colon cancer surgery in elderly patients
Yao ZHANG ; Songhua LIU ; Huimin WANG ; Liyan CAO ; Jiahui ZHAO ; Jie LI ; Wanting LUO ; Ling LI ; Lu LI ; Zhigang CHENG
Journal of Chinese Physician 2023;25(11):1624-1629
Objective:To investigate the effect of ultrasound-guided lumbar quadratus muscle plane block combined with general anesthesia on the analgesic effect during and after laparoscopic colon cancer radical surgery in elderly patients.Methods:A prospective study was conducted on 61 elderly patients who underwent radical colon cancer surgery at the Changsha Central Hospital Affiliated to South China University from May 2022 to February 2023, with American Society of Anesthesiologist (ASA) grades Ⅱ to Ⅲ. They were randomly divided into SA and GA groups using a random number table method, with 31 patients in the SA group and 30 patients in the GA group. The SA group received ultrasound-guided plane block of the upper lumbar quadratus muscle in the arcuate ligament combined with general anesthesia, while the GA group received simple general anesthesia. Record the mean artery pressure (MAP) and heart rate (HR) of two groups of patients at the time of entering the operating room (T 0), 5 minutes before skin incision (T 1), at skin incision (T 2), 30 minutes after surgery (T 3), at surgery end (T 4), and during anesthesia resuscitation and extubation (T 5); The Visual Analogue Scale (VAS) of two groups of patients at T 5, upon returning to the ward (T 6), 6 hours (T 7), 12 hours (T 8), 24 hours (T 9), and 48 hours (T 10) after surgery were recorded; The dosage of sufentanil, remifentanil, and propofol used during surgery, anesthesia recovery time, total number of analgesic pump presses within 48 hours after surgery, Lovett muscle strength score, early postoperative recovery, and adverse reactions within 48 hours after surgery were also recorded. Resultsl:The MAP and HR of the SA group were lower than those of the GA group at T 1-5 (all P<0.05); The VAS score of the SA group was lower than that of the GA group at T 5-10 (all P<0.05); The intraoperative dosage of propofol, remifentanil, and sufentanil in the SA group was lower than that in the GA group (all P<0.05); The anesthesia recovery time and first time out of bed in the SA group were earlier than those in the GA group (all P<0.05); The total number of times the analgesic pump was pressed within 48 hours after surgery was less than that of the GA group ( P<0.05); The incidence of nausea and vomiting within 48 hours after surgery in the SA group was lower than that in the GA group ( P<0.05). There was no statistically significant difference in postoperative Lovett muscle strength score, hospital stay, and dizziness incidence between the two groups of patients (all P>0.05). Conclusions:Ultrasound-guided lumbar quadratus muscle plane block on the arcuate ligament can significantly reduce the dosage of intraoperative general anesthesia drugs and postoperative analgesics in elderly patients undergoing laparoscopic colon cancer surgery, reduce postoperative pain scores, effectively alleviate postoperative pain, and thus advance the patient′s first time out of bed activity, reduce postoperative complications, and promote rapid recovery.
8.Study on the effect and mechanisms of SIGLEC-15 on laryngeal squamous cell carcinoma
Songhua LI ; Jin SUN ; Wenming CHEN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2022;57(6):724-731
Objective:To investigate the effect of sialic acid combined with immunoglobulin-like lectin 15 (SIGLEC-15) on laryngeal squamous cell carcinoma (LSCC) and underlying mechanisms.Methods:The Cancer Genome Atlas (TCGA), Cancer Cell Line Encyclopedia (CCLE) and Gene Expression Profiling Interactive Analysis (GEPIA2) databases were used for bioinformatics analysis. Cell Counting Kit-8 (CCK8), flow cytometry, and Transwell method were used respectively to detect proliferation, apoptosis, cell cycle, metastasis and invasion behaviors of the cells. Gene chip method was used for detecting up-regulated and down-regulated genes and performing enrichment analysis of differential genes. Western Blotting (WB) and Quantitative Real-time PCR (qPCR) were used to detect the expressions of proteins. Tumor formation experiments in nude mice were used to detect the effect of SIGLEC-15 on the growth of transplanted tumors. Wilcoxon rank sum test, t-test and Log-rank test were used for statistical analysis. Results:SIGLEC-15 was highly expressed in laryngeal squamous cell carcinoma and closely related to life in being. TU686 SIGLEC-15-with low expression of SIGLEC-15 was constructed. Compared to TU686 SIGLEC-15+, TU686 SIGLEC-15-showed significantly reduced activities of proliferation (48 h: 1.32±0.23 vs. 2.56±0.37, t=6.59, P<0.05), migration (1 036.52±51.22 vs. 1 819.62±180.24, t=7.22, P<0.05) and invasion (469.21±112.25 vs. 961.45±102.03, t=7.85, P<0.05); early increased apoptosis ((23.27±1.12)% vs. (5.64±1.61)%, t=11.32, P<0.05); blocked cell cycle at G0/G1 ((59.32±3.65)% vs. (35.46±3.57)%, t=9.85, P<0.05); the knockdown of SIGLEC-15 resulted in up-regulation of 864 genes, down-regulation of 357 genes, with significant changes in molecules of cell cycle, apoptosis and JAK/STAT signal pathways, and the expressions of p-JAK2, p-STAT3, Caspase-3, Bad, Bcl-2, and Cyclin d1 proteins. Tumor formation experiments in nude mice showed that at 8 weeks after the tumors were implanted, the growth transplanted tumors of TU686 SIGLEC-15-cell group was slower than that of TU686 SIGLEC-15+cell group, with significant difference in the mean tumor weights between two groups ((0.382±0.054) g vs. (1.277±0.126) g, t=8.44, P<0.05), while the expression of SIGLEC-15 was lower in the transplanted tumors of SIGLEC-15 knockdown group compared to control group, with significant difference(11.29±2.17 vs. 36.25±7.56, t=9.28, P<0.05). Conclusion:SIGLEC-15 is highly expressed in LSCC and can promote the progression of LSCC through the JAK2-STAT3 pathway.
9.Effect of ultrasound guided thoracic paravertebral nerve block on quality of recovery from general anesthesia in patients with tuberculous empyema surgery in post anesthesia recovery unit
Songhua LIU ; Yi FANG ; Liyan CAO ; Hongyi TAN ; Qiongcan LI ; Zhigang CHENG
Journal of Chinese Physician 2021;23(1):10-14
Objective:To study on the effect of ultrasound-guided thoracic paravertebral nerve (TPVB) block on quality of recovery from general anesthesia in tuberculosis patients with fiberboard exfoliation in post anesthesia recovery unit (PACU).Methods:From May 2018 to December 2019, 40 tuberculosis patients in Changsha Central Hospital with pulmonary fibreboard exfoliation and focal abscess lesions cleaning were randomly divided into two groups, with 20 patients in each group. The patients in group A received endobronchial general anesthesia and in group B received ultrasound-guided TPVB combined with endobronchial general anesthesia. Patients in the two groups were maintained under anesthesia by propofol, and the bispectral index (BIS) was maintained within the range of 40-50. The dosage of propofol and sufentanil was adjusted according to changes in BIS and hemodynamics. The mean arterial pressure (MAP), heart rate (HR) in two groups of patients were recorded at before anesthesia induction (T 0), before cutting leather (T 1), cut skin after (T 2), the end of operation (T 3), extubation time (T 4), and T 5 (time of leaving PACU). The visual analogue scale (VAS) of all patients in resting and cough state was recorded at 5, 30 min after extubation and the time of leaving PACU. The dosage of propofol and sufentanil in the operation and the additional dosage of sufentanil in PACU were recorded in both two groups. And the respiratory recovery time, consciousness recovery time, extubation time and sedation agitation scale(SAS) were observed. The adverse reactions such as nausea, vomiting, drowsiness and hypotension were observed in PACU. Results:Compared with group A, MAP and HR of patients at T 2, T 3, T 4, T 5 in group B were more stable during anesthesia, and VAS of patients in group B were lower than that in group A at each time point after extubation ( P<0.05). The dosage of sufentanil and propofol in group B were (35.92±8.12)μg and (749.56±95.30)mg respectively, which were significantly lower than those in group A [(45.74±4.42)μg and (862.83±105.34)mg, P<0.05]; the dosage of sufentanil in postoperative anesthesia recovery room of group B was (5.26±2.10)μg, significantly less than that of group A (10.35±5.86)μg ( P<0.05). The respiratory recovery time, consciousness recovery time and extubation time in group B were (12.92±5.12) min, (20.56±5.10) min and (26.87 ± 6.16) min, which were shorter than those in group A [(15.74±4.72)min, (25.83±5.34)min and (35.35±5.80)min, P<0.05]. The incidence of postoperative nausea, vomiting, lethargy and hypotension in group B were 10%, 10%, 35% and 20%, which were significantly lower than those in group A (30%, 20%, 75% and 45%, P<0.05). Conclusions:Ultrasound-guided paravertebral nerve block may significantly reduce the dosage of opioid analgesics for general anesthesia in tuberculosis patients with fiberboard exfoliation, accelerate the speed of anesthesia recovery, reduce the agitation during recovery, and improve the quality of anesthesia recovery.
10.RIP3-mediated necroptosis induced by radiation injury in neuronal cells
Songhua YANG ; Shixiong HUANG ; Biao ZENG ; Qian DONG ; Xiaocong ZHU ; Na ZENG ; Bin LI ; Guanzhi ZHOU ; Yifang CHEN ; Huiting YANG ; Jian LI ; Yingrui SHI
Chinese Journal of Radiation Oncology 2020;29(12):1124-1129
Objective:To observe the presence or absence of necroptosis in PC12 cells after radiation injury, and to detect the expression of receptor-interacting protein 3(RIP3) and evaluate its regulatory effect on necroptosis.Methods:PC12 cells were treated with different doses of irradiation and their necroptosis was detected by lactate dehydrogenase (LDH) release at different time points. After pretreatment with necroptosis inhibitor Necrostatin-1(Nec-1), the changes of cell necroptosis were detected by LDH. The expression level of RIP3 after irradiation intervention was detected by Western blot (WB). After pretreatment with the RIP3-specific inhibitor GSK′872, the changes of cell necroptosis were detected by LDH. The best transfection sequence of RIP3 knockout was screened by WB. The cells were divided into the control group, irradiation group, solvent control group, no-load control group and pretreatment group. WB, immunofluorescence staining, MTT, LDH and Annex V-fluorescein Isothiocyanate/Propidium Iodide (AnnexV-FITC/PI) flow cytometry were used for detection and analysis.Results:After 4 Gy irradiation, the degree of cell necrosis was the highest after 3 hours of culture, and the expression level of RIP3 protein was up-regulated. The cell necrosis was decreased after Nec-1, GSK′872 and RIP3 gene knockdown pretreatment.Conclusions:The radiation injury of 4 Gy can induce the necroptosis of PC12 cells, and the most significant effect can be observed when cultured for 3 hours after irradiation. RIP3 is involved in the process of necroptosis of PC12 cells induced by radiation injury, and plays a pivotal positive regulatory role.

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