1.A Case of EML4-ALK Fusion V1 Subtype Lung Adenocarcinoma Detected by RNA-based NGS
XU YUE ; MU NING ; LIU MEI ; WU SHENGNAN ; MA CHUNHUA
Chinese Journal of Lung Cancer 2024;27(6):480-484
Lung cancer is the malignant tumor with the highest incidence and mortality rate worldwide.For lung adenocarcinoma,identifying specific gene mutations,fusions,and giving corresponding targeted drugs can greatly improve the survival time of the patients.Among them,anaplastic lymphoma kinase(ALK)fusion occurs in 3%-7%of non-small cell lung cancer(NSCLC).In clinical practice,a variety of detection methods can be used to determine the ALK fusion status,but false negative test results are possible.This paper retrospectively analyzed the diagnosis and treatment of a patient with lung adeno-carcinoma,judged the ALK fusion status by various detection methods.Among them,immunohistochemistry(IHC)(Ventana D5F3),RNA based next-generation sequencing(RNA-based NGS)confirmed positive echinoderm microtubule associated protein like 4(EML4)-ALK fusion,while DNA-based NGS was negative.This paper analyzed the detection methods of ALK fusion,in order to clarify which detection method is the most accurate and simple to choose in different clinical cases and guide the subsequent treatment.
2.Effect of hydrogen on lipopolysaccharide and nigericin-induced pyroptosis in macrophages and the role of lncRNA NEAT1
Tao YANG ; Man LI ; Ling DING ; Shenyue PANG ; Lei WANG ; Licheng GENG ; Jiangang XU
Chinese Journal of Anesthesiology 2024;44(2):232-237
Objective:To evaluate the effect of hydrogen on lipopolysaccharide (LPS) and nigericin-induced pyroptosis in macrophages and the role of long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1).Methods:Human monocyte-derived macrophages THP-1 cells were cultured in vitro and divided into 4 groups ( n=25 each) using a random number table method: control group (group C), LPS and nigericin group (group LN), hydrogen-rich medium+ LPS and nigericin group (group H+ LN), and lentiviral transfection+ hydrogen-rich medium+ LPS-nigericin group (group LV+ H+ LN). THP-1 cells were cultured in the common culture medium for 24 h in group C. LPS at a final concentration of 100 ng/ml and nigericin 10 μmol/L were added to the culture medium, and the cells were incubated for 24 h in group LN. In group H+ LN, the culture medium was replaced with 0.6 mmol/L hydrogen-rich medium, then LPS at a final concentration of 100 ng/ml and nigericin 10 μmol/L were immediately added, and the cells were incubated for 24 h. In group LV+ H+ LN, THP-1 cells over-expressing NEAT1 stably after being transfected with lentivirus were used, then LPS at a final concentration of 100 ng/ml and nigericin 10 μmol/L were immediately added, and the cells were incubated for 24 h. Cell viability was detected by CCK-8 assay. Lactic dehydrogenase (LDH) release was assessed by colorimetric method. The amount of LDH released was measured by colorimetry. The concentrations of interleukin-1beta (IL-1β) and IL-18 in culture medium were measured by enzyme-linked immunosorbent assay. The pyroptotic rate was detected by flow cytometry. The expression of nucleotide-binding oligomerization domain-like receptor-containing pyrin domain 3 (NLRP3), apoptosis-associated speck-like protein (ASC), caspase-1 and gasdermin D (GSDMD) was detected by Western blot. The expression of NEAT1 gene was determined by quantitative real-time polymerase chain reaction. Results:Compared with group C, the cell viability was significantly decreased, the amount of LDH released, concentrations of IL-1β and IL-18, and pyroptotic rate were increased, and the expression of NEAT1 gene, NLRP3, ASC, caspase-1 and GSDMD was up-regulated in group LN ( P<0.05). Compared with group LN, the cell viability was significantly increased, the amount of LDH released, concentrations of IL-1β and IL-18, and pyroptotic rate were decreased, and the expression of NEAT1 gene, NLRP3, ASC, caspase-1 and GSDMD was down-regulated in group H+ LN ( P<0.05). Compared with group H+ LN, the cell viability was significantly decreased, the amount of LDH released, concentrations of IL-1β and IL-18, and pyroptotic rate were increased, and the expression of NEAT1 gene, NLRP3, ASC, caspase-1 and GSDMD was up-regulated in group LV+ H+ LN ( P<0.05). Conclusions:Hydrogen can ameliorate LPS and nigericin-induced pyroptosis in macrophages, and the mechanism may be associated with down-regulating the expression of lncRNA NEAT1.
3.Analysis of advanced fibrosis in metabolic dysfunction-associated fatty liver disease patients with chronic hepatitis B
Xue WU ; Ping LI ; Yuqiang MI
Chinese Journal of Internal Medicine 2024;63(1):53-58
Objective:To investigate the clinical and pathological characteristics of chronic hepatitis B (CHB) with metabolic dysfunction-associated fatty liver disease (MAFLD), as well as associations with advanced fibrosis.Methods:CHB patients who underwent liver biopsy at Tianjin Second People′s Hospital from June 2016 to September 2019 were included in the study. The patients were divided into two groups based on whether they had concomitant MAFLD; a CHB group and a MAFLD-CHB group. t-tests and Chi-square tests were used to compare pathological characteristics and basic features in the two groups. Logistic regression analysis was used to analyze factors associated with advanced fibrosis. Results:The CHB group included 110 patients, and the MAFLD-CHB group included 272 patients. There were significant differences in smoking, alcohol consumption, hypertension incidence, body metabolic index, alanine aminotransferase, gamma-glutamyl transferase (GGT), high-density lipoprotein, low-density lipoprotein, fasting plasma glucose, and platelets (PLT) between the two groups (all P<0.05). The MAFLD-CHB group had a higher incidence of advanced fibrosis than the CHB group ( P<0.05). In logistic regression analysis MAFLD [odds ratio ( OR)=2.204, 95% confidence interval ( CI) 1.018-4.774, P=0.045], GGT ( OR=1.008, 95% CI 1.002-1.013, P=0.005), and PLT ( OR=0.995, 95% CI 0.991-0.999, P=0.019) were associated with advanced fibrosis (all P<0.05). In the MAFLD-CHB group type 2 diabetes ( OR=3.281, 95% CI 1.375-7.832, P=0.007), GGT ( OR=1.011, 95% CI 1.003-1.018, P=0.005), and PLT ( OR=0.993, 95% CI 0.988-0.998, P=0.004) were associated with advanced fibrosis ( P<0.05). Conclusion:Patients with MAFLD-CHB are more likely to develop advanced fibrosis than patients with CHB alone. In the MAFLD-CHB group type 2 diabetes mellitus was associated with advanced fibrosis. It is important to strictly control relevant risk factors in MAFLD-CHB patients, especially in patients with type 2 diabetes.
4.Research progress on antitumor activity and mechanism of solasonine
Chinese Journal of Cancer Biotherapy 2024;31(4):404-409
[摘 要] 澳洲茄碱(SS)是一种天然来源的生物活性小分子化合物,已经被证实具有抗菌、抗炎和抗肿瘤等功效。在抗肿瘤作用方面,SS可以通过多种机制在不同类型肿瘤中有效发挥抗肿瘤作用,其作用机制包括诱导肿瘤细胞凋亡、阻滞肿瘤细胞周期、诱导肿瘤细胞铁死亡、调控肿瘤相关非编码RNA、调节免疫和炎症反应、削弱肿瘤细胞的侵袭和转移能力、抑制糖酵解进程和肿瘤干细胞的产生等,涵盖了肺癌、肝癌、胃癌、乳腺癌、胆管癌、膀胱癌、胰腺癌、白血病、骨肉瘤等多种癌症类型。阐明SS在不同类型肿瘤中的抗肿瘤活性及其作用机制,为促进SS抗肿瘤作用机制的进一步研究和开发更有效安全的抗肿瘤药物提供了重要的理论基础。
5.Effects of intravenous and topical tranexamic acid on blood loss and inflammatory response after posterior cervical decompressive laminectomy
Chinese Journal of Tissue Engineering Research 2024;28(21):3367-3372
BACKGROUND:With the increase of the incidence of cervical spondylosis,it is particularly important to control the postoperative bleeding and related inflammatory reaction of cervical vertebra. OBJECTIVE:A prospective study was conducted to investigate the combined effect of intravenous and topical tranexamic acid of different concentrations on postoperative blood loss and inflammatory response during posterior cervical decompressive laminectomy combined with lateral mass screw internal fixation and bone graft fusion. METHODS:From January 2020 to December 2022,150 patients who were scheduled to undergo posterior cervical decompressive laminectomy combined with lateral mass screw internal fixation and bone graft fusion for cervical spondylotic myelopathy in Tianjin People's Hospital were enrolled in the study.Patients were divided into A,B,and C groups(n=50)by randomized double-blind lottery.The patients in the A,B,and C groups were treated with 1%,3%,and 5%intravenous and topical tranexamic acid solution,respectively.Postoperative blood loss and inflammation-related indicators were compared among three groups.The occurrence of venous thromboembolism and hematoma was observed after operation. RESULTS AND CONCLUSION:(1)There were significant differences in blood loss-related indexes,such as intraoperative blood loss,overall blood loss,and occult blood loss among the three groups(P<0.01).The above indexes were significantly lower in groups B and C than that in group A.There was no significant difference between groups B and C(P>0.05).(2)On days 1 and 3 after surgery,the levels of inflammatory indicators including serum C-reactive protein and interleukin-6 were increased to varying degrees in the three groups compared to before surgery(P<0.05).As the drug concentration increased,its expression decreased,and there was a significant difference among the three groups(P<0.001).(3)Two cases of intramuscular venous thrombosis appeared in each of the three groups after surgery.No hematoma or pulmonary embolism occurred in all three groups.(4)It is concluded that in posterior cervical decompressive laminectomy combined with lateral mass screw internal fixation and bone graft fusion,intravenous combined with topical application of different concentrations of tranexamic acid is effective in reducing perioperative blood loss and inflammatory response.3%tranexamic acid achieves the same effect without significantly increasing the risk of postoperative venous thromboembolism.
6.Correlation between sagittal alignment and clinical symptom improvement after lumbar fusion
Zhao FANG ; Pan QIAO ; Tiantong XU
Chinese Journal of Tissue Engineering Research 2024;28(24):3778-3782
BACKGROUND:Up to date,there is seldom research about the correlation between the harmonious alignment degree of lumbar lordosis and pelvic incidence and the improvement of postoperative clinical symptoms in patients with lumbar stenosis fusion. OBJECTIVE:To investigate the harmonious alignment degree of lumbar lordosis and pelvic incidence in patients with lumbar spinal stenosis and fusion,and compare the differences of spinal and pelvic sagittal parameters and clinical outcomes among different harmonious alignment degrees. METHODS:The medical records of 110 patients with lumbar spinal stenosis(48 males and 62 females,aged 57±11 years)who were treated with posterior lumbar laminar decompression and internal fixation in the Department of Spinal Surgery,Tianjin Union Medical Center from December 2019 to December 2021 were retrospectively analyzed.The postoperative follow-up was more than 1 year.According to the harmonious alignment degree of lumbar lordosis and pelvic incidence,they were divided into two groups:Group A(n=47)(pelvic incidence-lumbar lordosis>10°)and group B(n=63)(pelvic incidence-lumbar lordosis≤10°).The changes of clinical scores(low back pain visual analog scale score and Oswestry disability index)and spinal and pelvic sagittal parameters before and after surgery were compared between the two groups.The correlation between clinical symptom improvement and spinal and pelvic imaging parameters was analyzed. RESULTS AND CONCLUSION:(1)There were no significant differences in preoperative pelvic sagittal parameters and clinical function between the two groups(P>0.05).(2)There were no significant differences in operation time and blood loss between the two groups(P>0.05).(3)At the last postoperative follow-up,the visual analog scale score and Oswestry disability index in group B were significantly improved compared with group A(P=0.000,P=0.005).Lumbar lordosis and sacral slope were also significantly improved(P=0.000,P<0.05),and pelvic tilt was more significantly recovered(P=0.000).(4)Pearson correlation coefficient method results showed that the improvement of visual analog scale score of low back pain was significantly correlated with the improvement of pelvic tilt(r=0.209,P=0.008),lumbar lordosis(r=-0.133,P=0.021)and sacral slope(r=-0.156,P=0.031).Pelvic tilt was significantly correlated with Oswestry disability index(r=0.196,P=0.014).(5)It is concluded that reconstructed lumbar lordosis and reduced pelvic tilt for the harmonious alignment(pelvic incidence-lumbar lordosis≤10°)can effectively improve the clinical symptoms of patients.
7.Influence of preoperative C7/T1 foraminal area on the efficacy of posterior cervical laminoplasty in the treatment of cervical spondylotic myelopathy
Lilong ZHANG ; Rui SHAO ; Yannan GENG
Chinese Journal of Spine and Spinal Cord 2024;34(5):458-462
Objectives:To investigate the effects on the efficacy of posterior cervical laminoplasty in patients with cervical spondylotic myelopathy of different C7/T1 foraminal areas before surgery.Methods:76 patients who underwent posterior cervical open-door expansive laminoplasty for cervical spondylotic myelopathy in our hospital from September 2021 to September 2022 were analyzed retrospectively,including 58 males and 18 females,aged 64.4±8.5 years old.The area of C7/T1 foramina of patients was measured on the double oblique X-ray images before operation,and the patients were divided into two groups on the basis of the av-erage C7/T1 foraminal area:Group A,C7/T1 foraminal area ≤average value(40 patients),and group B,C7/T1 foraminal area>average value(36 patients).The operative time and intraoperative bleeding were collected and compared between groups,and the Japanese Orthopaedic Association(JOA)scores before surgery,3 months af-ter surgery,and 12 months after surgery were obtained to calculate the JOA score improvement rate;The axi-al symptoms at 12 months after surgery were recorded,and T test,analysis of variance,and chisquare test were used to analyze whether different preoperative C7/T1 forminal areas of patients affected the efficacies after posterior cervical laminoplasty.Results:The foraminal areas of C7/T1 was 35.2±9.7mm2 in group A and 65.7±13.1mm2 in group B,and C2-C7 Cobb angle before operation was 14.0°±3.6° in group A and 16.0°±5.5° in group B,with statistical differences respectively(P<0.05).Group A was not significantly different from group B in terms of intraoperative bleeding(176.8±88.2mL vs 183.6±100.2mL)and operative time(127.5±23.6min vs 120.3±32.6min)(P>0.05).The JOA scores of group A and group B were 10.9±2.0 and 10.3±2.1 before operation,without statistical difference(P>0.05);The JOA scores of group A and group B were 12.8±1.5 and 14.0±2.2 at postoperative 3 months and 14.1±1.5 and 15.9±1.7 at 12 months after operation,with statistical differences respectively(P<0.05).There were statistical differences in the improvement rates of JOA scores between the two groups at postoperative 3 months and 12 months,respectively(P<0.05).The incidence of axial symptoms 12 months after operation in group A and group B was 42.5%and 19.4%,respectively,with statistical difference(P<0.05).Conclusions:Patients with larger C7/T1 foraminal area have better postoperative neurological recovery,higher rate of JOA improvement,and lower incidence of postoperative axial symptoms.
8.The significance of lumbosacral curve vertebral body leveling in coronal correction of degenerative scoliosis
Chuanxu LI ; Ziyang LIU ; Xuanhao FU
Chinese Journal of Spine and Spinal Cord 2024;34(10):1030-1037
Objectives:To analyze the correction of postoperative coronal imbalance in adult degenerative scoliosis patients undergoing spinal fusion surgery,and to explore the impact of leveling of the most tilted vertebra(L4 or L5)in the lumbosacral curve on the correction of coronal imbalance.Methods:A retrospective analysis was conducted on 61 patients(17 male and 44 female;aged 51 to 78 years,mean 63.57±8.41 years;body mass index 15.79-35.55kg/m2,mean 25.83±3.69kg/m2)who underwent degenerative scoliosis correction surgery between January 2019 and March 2023 in Tianjin Union Medical Center.The patients were catego-rized based on the Obeid classification and the length of fusion segments into concave long segment group(17 cases),concave short segment group(15 cases),convex long segment group(15 cases),and convex short seg-ment group(14 cases).Coronal radiographic parameters[Cobb angle,coronal balance distance(CBD),coronal tilt angle]and sagittal radiographic parameters(sagittal vertical axis,thoracic kyphosis,lumbar lordosis,pelvic inci-dence,pelvic tilt,sacral slope)were measured on full spine anteroposterior and lateral X-rays preoperatively and on 3d postoperatively.The postoperative coronal and sagittal imbalance corrections were compared be-tween groups.Pearson correlation analysis was performed to investigate the relationship between the correction ratio of the maximal coronal tilt angle in the lumbosacral curve(L4 or L5)and postoperative CBD.The opera-tive time,blood loss,and visual analogue scale(VAS)pre-and postoperatively were compared between the four groups.Results:The postoperative coronal Cobb angle,CBD,and maximal coronal tilt angle at L4 or L5 of the concave long segment group,convex long segment group,and convex short segment group were statisti-cally different from those before operation(P<0.05).In the concave short segment group,the postoperative coronal Cobb angle and maximal coronal tilt angle at L4 or L5 were significantly different from those before operation(P<0.05),while no significant difference was found between pre-and postoperative CBD(P=0.085).Pearson correlation analysis revealed a significant negative correlation(r=-0.570,P<0.01)between postoperative CBD and coronal tilt correction ratio in the convex group,with the linear regression equation being postoper-ative CBD=32.81-28.38 x coronal tilt correction ratio.When the correction ratio of the maximal coronal tilt angle at L4 or L5 exceeding 45%in convex coronal malalignment patients,it's predicted that the CBD could be corrected to within 20mm.No significant correlation was found between postoperative CBD and the coronal tilt correction ratio in the concave group(r=0.098,P=0.59).Patients in both the convex and concave short segment groups had shorter operative time and less blood loss than long segment groups.Postoperative VAS scores improved significantly in all groups(P<0.05).Conclusions:In adult degenerative scoliosis patients with convex coronal malalignment,both short and long segment fusions are capable of correcting coronal imbalance.Concave coronal malalignment patients require long segment fusion for the correction of coronal imbalance.To achieve optimal coronal balance in convex coronal malalignment patients,a correction ratio of the maximal coronal tilt angle at L4 or L5 shall exceed 45%.
9.Application of imaging mass cytometry in gastroenterological carcinoma
Zehan LIU ; Jing XUN ; Aimin ZHANG ; Botao WANG ; Lanqiu ZHANG ; Ximo WANG ; Qi ZHANG
Chinese Journal of Immunology 2024;40(4):866-871
Imaging mass cytometry(IMC)is a new technology integrating mass spectrometry,high-resolution laser ablation and immunohistochemistry/cytochemistry.With a unique high-dimensional perspective to comprehensively and accurately depict the complex phenotypes,signaling pathways,and tumor immune interactions in the tissue and tumor microenvironment,is widely used in gastrointestinal tumors.This article reviews the application of IMC in depicting the panorama of tumor microenvironment,revealing tu-mor spatial heterogeneity,clarifying tumor pharmacological mechanism,assisting new drug development,and evaluating the efficacy of immunotherapy dynamically in digestive system tumors.
10.Effects of miR-17-5p targeting B7-H3 on biological characteristics and immunomodulatory effects of colorectal cancer cells
Yang HUA ; Xiukun MA ; Zhu HONG ; Xianglong LIU
Chinese Journal of Immunology 2024;40(7):1431-1435,1440
Objective:To investigate the regulatory role of miR-17-5p on B7-H3 and its immunomodulatory role in the develop-ment of colorectal cancer.Methods:Twenty-five pairs of human colorectal cancer tissues and adjacent normal tissues were collected,and the expression levels of miR-17-5p,B7-H3 and PD-L1 were analyzed by real-time fluorescence quantitative PCR,protein immuno-blotting and immunohistochemical assays.Human colorectal cancer HCT-116 cells were divided into four groups:miR-NC group,miR-17-5p mimic group,si-NC group and si-B7-H3 group.The growth,invasive ability and apoptosis rate of HCT-116 cells were ana-lyzed by cell colony formation assay,Transwell assay and flow cytometry.The regulatory effect of miR-17-5p on B7-H3 was analyzed by luciferase reporter gene assay.The levels of IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α cytokines in the cell culture medium were analyzed by ELISA.Results:Compared with paracancerous tissues,miR-17-5p expression was decreased and B7-H3 expression was increased in colorectal cancer tissues.miR-17-5p mimic and si-B7-H3 were able to reduce the number of HCT-116 cell colony forma-tion and invasion,promote HCT-116 cell apoptosis,and decrease the levels of IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α cytokines in culture medium.In addition,miR-17-5p mimic and si B7-H3 were able to inhibit PD-L1 expression in HCT-116 cells.Conclusion:miR-17-5p can targeting inhibit B7-H3 expression,affect the immunomodulatory role in colorectal cancer development,and inhibit metastasis and promote apoptosis of colorectal cancer cells.

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