1.Effects of long non-coding RNA KIAA0125 on proliferation and apoptosis of acute myeloid leukemia U937 cells
Huali HU ; Fahua DENG ; Yuancheng LIU ; Siqi WANG ; Jingxin ZHANG ; Tingting LU ; Hai HUANG ; Sixi WEI
Chinese Journal of Tissue Engineering Research 2025;29(19):3983-3991
BACKGROUND:U937 cells can be used as a cell model for studying the biological characteristics,signaling pathways,and therapeutic targets of acute myeloid leukemia.Although it has been reported that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia,its biological function in U937 cells remains unclear,and its mechanism of action in the occurrence and development of acute myeloid leukemia needs to be further clarified. OBJECTIVE:To investigate the expression level of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia and its effect on the proliferation and apoptosis of U937 cells. METHODS:RNA-sequencing was used to analyze the bone marrow monocyte samples from acute myeloid leukemia patients,and the differentially expressed gene long non-coding RNA KIAA0125 was screened.The expression of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia was detected by qRT-PCR.The relationship between long non-coding RNA KIAA0125 mRNA expression and prognosis in bone marrow cells of 173 acute myeloid leukemia patients and 70 healthy people was statistically analyzed by GEPIA database.Subsequently,recombinant lentivirus technology and CRISPR/Cas9-SAM technology were used to construct U937 cell lines with knockdown/overexpression of long non-coding RNA KIAA0125.qRT-PCR was used to detect the knockdown/overexpression efficiency of long non-coding RNA KIAA0125.Next,CCK-8 assay,flow cytometry,and western blot assay were used to detect the effects of knockdown/overexpression of long non-coding RNA KIAA0125 on the proliferation and apoptosis of U937 cells.Finally,western blot assay was used to detect the effect of knockdown/overexpressed long non-coding RNA KIAA0125 on Wnt/β-catenin signaling pathway-related proteins. RESULTS AND CONCLUSION:(1)The results of qRT-PCR showed that long non-coding RNA KIAA0125 was highly expressed in peripheral blood of acute myeloid leukemia patients.The results of GEPIA database showed that long non-coding RNA KIAA0125 was highly expressed in bone marrow cells of acute myeloid leukemia patients,and the high expression group had worse overall survival.(2)The knockdown efficiency of long non-coding RNA KIAA0125 in knockdown group was 70%,and the U937 cells that stably down-regulated long non-coding RNA KIAA0125 expression were successfully constructed.The expression of long non-coding RNA KIAA0125 in overexpression group was four times that of vector group,and stable U937 cells were successfully constructed.(3)Knockdown of long non-coding RNA KIAA0125 inhibited the proliferation of U937 cells and promoted their apoptosis.Overexpression of long non-coding RNA KIAA0125 promoted the proliferation of U937 cells but had no significant effect on the apoptosis of U937 cells.(4)Knockdown of long non-coding RNA KIAA0125 inhibited the activity of Wnt/β-catenin signaling pathway,while overexpression of long non-coding RNA KIAA0125 activated Wnt/β-catenin signaling pathway.These results confirm that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia peripheral blood.Long non-coding RNA KIAA0125 may affect the proliferation and apoptosis of U937 cells by regulating the Wnt/β-catenin signaling pathway,and may be a potential prognostic marker for acute myeloid leukemia.
2.Effect of Zuogui Jiangtang Tongmai Prescription on Inflammatory Injury of Human Umbilical Vein Endothelial Cells Induced by High Glucose and LPS Based on GPR43/β-arrestin-2/IκBα/NF-κB Pathway
Lanyu PENG ; Jingxin YAO ; Yujia LI ; Dingxiang LI ; Xun LIU ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):64-74
ObjectiveTo investigate the effects and mechanism of Zuogui Jiangtang Tongmai prescription (ZJTP) on human umbilical vein endothelial cells (HUVECs) damaged by high glucose combined with lipopolysaccharide (LPS). MethodThe survival rate of cells was determined by cell counting kit-8 (CCK-8), and the level of tumor necrosis factor-α (TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA) to determine the optimal injury concentration and action time of LPS, as well as the optimal action concentration of ZJTP drug-containing serum. HUVECs were divided into a blank control group, a model group, a ZJTP drug-containing serum group, and an SCFA mixed liquid group. ELISA was used to detect the level of endothelin-1 (ET-1), nitric oxide (NO), interleukin-1β (IL-1β), interleukin-6 (IL-6), and TNF-α. Western blot was performed to detect the protein expression of G protein-coupled receptor43 (GPR43), β-suppressor protein-2 (β-arrestin-2), nuclear factor-κB suppressor α (IκBα), and nuclear factor κB p65 (NF-κB p65). The nucleation of NF-κB p65 was observed by immunofluorescence staining (IF). The role of GPR43 in the regulation of inflammatory injury was observed by means of small interfering ribonucleic acid (siRNA). The cells after intervention were divided into an empty carrier group, a ZJTP drug-containing serum group, a Si-GPR43 group, and a Si-GPR43 + ZJTP drug-containing serum group. The content of IL-1β, IL-6, and TNF-α was detected by ELISA. The protein expression of pathways was detected by Western blot. IF was used to observe the nucleation of NF-κB p65. ResultThe optimal molding condition was 1 mg·L-1 LPS for 24 h. The optimal drug intervention condition was 5% ZJTP drug-containing serum for 24 h. Compared with the blank control group, the content of ET-1 in the model group was significantly increased, and the content of NO was significantly decreased (P<0.01). The levels of inflammatory factors were significantly increased (P<0.01). The expressions of GPR43 and IκBα were significantly decreased, while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly increased (P<0.01). NF-κB p65 protein was transferred from the extranuclear to the intranuclear (P<0.01). Compared with the model group, the content of ET-1 in the ZJTP drug-containing serum group was decreased, and the content of NO was increased (P<0.05). The levels of inflammatory factors decreased (P<0.05). The protein expressions of GPR43 and IκBα were increased, while the expressions of β-arrestin-2 and NF-κB p65 were decreased (P<0.05). The amount of NF-κB p65 transferred from the intranuclear to the extranuclear decreased (P<0.01). The mechanism study showed that compared with the Si-GPR43 group, the content of IL-1β, IL-6, and TNF-α were significantly decreased after treatment with ZJTP drug-containing serum (P<0.01). The protein expressions of GPR43 and IκBα were significantly increased (P<0.01), while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly decreased (P<0.01). The amount of NF-κB p65 transferred from the extranuclear to the intranuclear decreased (P<0.01). ConclusionZJTP has a protective effect on HUVECs with high glucose and LPS-induced inflammatory injury, which may be related to the regulation of GPR43/β-arrestin-2/IκBα/NF-κB pathway.
3.SLAMF7 predicts prognosis and correlates with immune infiltration in serous ovarian carcinoma
Yalong DENG ; Lu ZHANG ; Changyuan DAI ; Yan XU ; Qiyu GAN ; Jingxin CHENG
Journal of Gynecologic Oncology 2024;35(6):e79-
Objective:
Signaling lymphocytic activation molecule family members (SLAMFs) play a critical role in immune regulation of malignancies. This study aims to investigate the prognostic value and function of SLAMFs in ovarian cancer (OC).
Methods:
The expression analysis of SLAMFs was conducted based on The Cancer Genome Atlas Ovarian Cancer Collection (TCGA-OV) and Gene Expression Omnibus (GEO) databases. Immunohistochemistry (IHC) was further performed on tissue arrays (n=98) to determine the expression of SLAMF7. Kaplan-Meier plotter and multivariate Cox regression model were used to evaluate the correlation of SLAMF7 expression with survival outcomes of patients. The molecular function of SLAMF7 in OC was further investigated using Gene Set Enrichment Analysis (GSEA).
Results:
SLAMF7 mRNA expression were significantly upregulated in OC tumor tissue compared to normal tissue. IHC revealed that SLAMF7 expression was located in the interstitial parts of tumor tissue, and higher SLAMF7 expression was associated with favorable survival outcomes. GSEA demonstrated that SLAMF7 is involved immune-related pathways. Further analysis showed that SLAMF7 had a strong correlation with the T cellspecific biomarker (CD3) but not with the B cell (CD19, CD22, and CD23) and natural killer cell-specific biomarkers (CD85C, CD336, and CD337). Furthermore, IHC analysis confirmed that SLAMF7 was expressed in tumor-infiltrating T cells, and the IHC score of SLAMF7 was positively correlated with CD3 (r=0.85, p<0.001).
Conclusion
SLAMF7 is expressed in the interstitial components of clinical OC tissue, and higher SLAMF7 expression indicated a favorable prognosis for patients with OC.Additionally, SLAMF7 is involved in T-cell immune infiltration in OC.
4.SLAMF7 predicts prognosis and correlates with immune infiltration in serous ovarian carcinoma
Yalong DENG ; Lu ZHANG ; Changyuan DAI ; Yan XU ; Qiyu GAN ; Jingxin CHENG
Journal of Gynecologic Oncology 2024;35(6):e79-
Objective:
Signaling lymphocytic activation molecule family members (SLAMFs) play a critical role in immune regulation of malignancies. This study aims to investigate the prognostic value and function of SLAMFs in ovarian cancer (OC).
Methods:
The expression analysis of SLAMFs was conducted based on The Cancer Genome Atlas Ovarian Cancer Collection (TCGA-OV) and Gene Expression Omnibus (GEO) databases. Immunohistochemistry (IHC) was further performed on tissue arrays (n=98) to determine the expression of SLAMF7. Kaplan-Meier plotter and multivariate Cox regression model were used to evaluate the correlation of SLAMF7 expression with survival outcomes of patients. The molecular function of SLAMF7 in OC was further investigated using Gene Set Enrichment Analysis (GSEA).
Results:
SLAMF7 mRNA expression were significantly upregulated in OC tumor tissue compared to normal tissue. IHC revealed that SLAMF7 expression was located in the interstitial parts of tumor tissue, and higher SLAMF7 expression was associated with favorable survival outcomes. GSEA demonstrated that SLAMF7 is involved immune-related pathways. Further analysis showed that SLAMF7 had a strong correlation with the T cellspecific biomarker (CD3) but not with the B cell (CD19, CD22, and CD23) and natural killer cell-specific biomarkers (CD85C, CD336, and CD337). Furthermore, IHC analysis confirmed that SLAMF7 was expressed in tumor-infiltrating T cells, and the IHC score of SLAMF7 was positively correlated with CD3 (r=0.85, p<0.001).
Conclusion
SLAMF7 is expressed in the interstitial components of clinical OC tissue, and higher SLAMF7 expression indicated a favorable prognosis for patients with OC.Additionally, SLAMF7 is involved in T-cell immune infiltration in OC.
5.SLAMF7 predicts prognosis and correlates with immune infiltration in serous ovarian carcinoma
Yalong DENG ; Lu ZHANG ; Changyuan DAI ; Yan XU ; Qiyu GAN ; Jingxin CHENG
Journal of Gynecologic Oncology 2024;35(6):e79-
Objective:
Signaling lymphocytic activation molecule family members (SLAMFs) play a critical role in immune regulation of malignancies. This study aims to investigate the prognostic value and function of SLAMFs in ovarian cancer (OC).
Methods:
The expression analysis of SLAMFs was conducted based on The Cancer Genome Atlas Ovarian Cancer Collection (TCGA-OV) and Gene Expression Omnibus (GEO) databases. Immunohistochemistry (IHC) was further performed on tissue arrays (n=98) to determine the expression of SLAMF7. Kaplan-Meier plotter and multivariate Cox regression model were used to evaluate the correlation of SLAMF7 expression with survival outcomes of patients. The molecular function of SLAMF7 in OC was further investigated using Gene Set Enrichment Analysis (GSEA).
Results:
SLAMF7 mRNA expression were significantly upregulated in OC tumor tissue compared to normal tissue. IHC revealed that SLAMF7 expression was located in the interstitial parts of tumor tissue, and higher SLAMF7 expression was associated with favorable survival outcomes. GSEA demonstrated that SLAMF7 is involved immune-related pathways. Further analysis showed that SLAMF7 had a strong correlation with the T cellspecific biomarker (CD3) but not with the B cell (CD19, CD22, and CD23) and natural killer cell-specific biomarkers (CD85C, CD336, and CD337). Furthermore, IHC analysis confirmed that SLAMF7 was expressed in tumor-infiltrating T cells, and the IHC score of SLAMF7 was positively correlated with CD3 (r=0.85, p<0.001).
Conclusion
SLAMF7 is expressed in the interstitial components of clinical OC tissue, and higher SLAMF7 expression indicated a favorable prognosis for patients with OC.Additionally, SLAMF7 is involved in T-cell immune infiltration in OC.
6.Role of intrahepatic microenvironment induced-autophagy in nonalcoholic fatty liver disease
Xuanxuan KOU ; Hua ZHANG ; Jingxin DENG ; Jiangang ZHANG
Journal of Clinical Hepatology 2023;39(6):1440-1445
Nonalcoholic fatty liver disease (NAFLD) is a series of abnormal liver lesions mainly characterized by excessive lipid deposition in hepatocytes, and it is also the most common chronic liver disease worldwide. Autophagy is a basic cellular process in which cells degrade their own components and participate in the maintenance of organ function and body homeostasis, and it is closely associated with the progression of NAFLD. High fat, hypoxia, and stress in human body may cause abnormal changes in extracellular microenvironment in the liver, and such abnormal microenvironment may promote the development and progression of NAFLD by inducing liver cell autophagy. This article reviews the role and mechanism of autophagy of liver cells such as hepatocytes, Kupffer cells, and hepatic stellate cells in the progression of NAFLD based on various microenvironment characteristics in the liver.
7.Clinical efficacy and safety of ex-vivo liver resection and liver auto-transplantation: a bayesian single-arm Meta-analysis
Yichen FAN ; Manjun DENG ; Jingxin YAN ; Li REN ; Haijiu WANG ; Haining FAN ; Qian LU
Chinese Journal of Hepatobiliary Surgery 2022;28(7):547-554
Objective:To evaluate the clinical efficacy and safety of ex vivo liver resection and autotransplantation (ELRA) by using a Bayesian single-arm Meta-analysis.Methods:Databases of PubMed, Embase, Cochrane Library, Web of Science, CNKI, and Wanfang were searched from January 1, 1990 to December 30, 2021 on ELRA studies. The Bayesian one-arm Meta-analysis was performed by using the statistical software of R (V4.1.2) and the Markov chain-Monte Carlo method was used to simulate the posterior distribution. The mortality rate within 30 days after operation, 1-year survival rate, major postoperative complications, R 0 resection rate and other related indexes were analyzed. Results:A total of 20 studies with 436 patients were included. Bayesian single-arm Meta-analysis showed that the 1-year survival rate after ELRA was 83.24% [95% highest posterior density ( HPD): 72.40%-92.05%]. The 1-year survival rates after surgery were 88.66% (95% HPD: 81.52%-94.50%) for patients with hepatic alveolar echinococcosis and 61.29% (95% HPD: 38.53%-93.68%) for patients with hepatic malignancies, respectively. The mortality rate within 30 d after surgery, the incidence of significant postoperative complications, and the R 0 resection rate were 6.96% (95% HPD: 4.47%-10.15%), 27.91% (95% HPD: 19.00%-38.30%), and 99.84% (95% HPD: 37.61%-100.00%), respectively. Renal failure was the most frequent cause of death after ELRA. Conclusion:ELRA is indicated for hepatic malignancies and hepatic alveolar echinococcosis when intrahepatic resection cannot be accomplished in vivo. The greatest benefit is observed in patients with hepatic alveolar echinococcosis, while only some patients with hepatic malignancies can benefit. The indications for ELRA for hepatic malignancies need to be further studied to define the subgroup of patients who can benefit from this operation.
8.Oncologic outcomes of early stage cervical cancer performed operation by different laparoscopic surgical procedures: analysis of clinical data from mutiple centers
Kaijian LING ; Yanzhou WANG ; Hui ZHANG ; Xuyin ZHANG ; Junjun YANG ; Chengyan LUO ; Bin SONG ; Wenxi ZHANG ; Li DENG ; Gongli CHEN ; Yudi LI ; Qunying HU ; Yong CHEN ; Xin WANG ; Jun ZHANG ; Jingxin DING ; Tong REN ; Shan KANG ; Keqin HUA ; Yang XIANG ; Wenjun CHENG ; Zhiqing LIANG
Chinese Journal of Obstetrics and Gynecology 2020;55(9):617-623
Objective:To evaluate the oncologic outcomes of different laparoscopic radical hysterectomy.Methods:From January 2011 to December 2014, the laparoscopic operation cases of cervical cancer at stage Ⅰb1, Ⅰb2, Ⅱa1 and Ⅱa2, including the histologic subtypes of squamous-cell carcinoma, adenocarcinoma and adenosquamous carcinoma, were collected in five clinical centers. The data were divided into two groups according to the surgical procedures, that is, modified laparoscopic-vaginal radical hysterectomy (mLVRH) and total laparoscopic radical hysterectomy (TLRH). The overall survival rate (OS), disease-free survival rate (DFS) at 5 years were retrospectively analyzed in this study.Results:There were 674 cases in total, including 377 cases of mLVRH, 297 cases of TLRH. (1) The OS at 5 years: the mLVRH was 96.1% and the TLRH was 92.0%, and the mLVRH was higher than that of TLRH ( P=0.010). Stratify analysis, including stage of disease (Ⅰb1 and Ⅱa1), histologic subtypes (squamous-cell carcinoma, adenocarcinoma), lymph node metastasis, revealed that, ① Stage of disease: in stage Ⅰb1, the OS at five years of mLVRH was higher than that in TLRH group (98.6% vs 93.6%, P=0.012). In stage Ⅱa1, there was significant difference between the two groups, the OS at five years of mLVRH and TLRH were 93.6% and 77.6% ( P=0.007). ② Histologic subtypes: for the OS at five years of squamous-cell carcinoma, mLVRH and TLRH were 96.1% and 92.3%, and there was significant difference ( P=0.046); for adenocarcinoma, the OS at five years were 91.0% and 88.6%, and there was no difference between two groups ( P=0.230). ③ Lymph node metastasis: the mLVRH and TLRH with lymph node metastasis, the OS at five years were 98.6% and 96.4%; the mLVRH and TLRH without lymph node metastasis, the OS at five years were 89.3% and 80.8%. There were no significant differences between the two groups,respectively ( P=0.156, P=0.093). (2) The DFS at 5 years: there was no significant difference between mLVRH and TLRH (94.1% vs 90.9%, P=0.220). Stratify analysis for stage of disease, the mLVRH group was higher than that in the TLRH group in stage Ⅰb1 (97.0% vs 92.8%, P=0.039). However, for stage Ⅱa1, there was no significant difference between mLVRH and TLRH group (88.2% vs 75.8%, P=0.074). Conclusions:The results of this retrospective study indicated that different laparoscopy surgical procedures had diverse oncologic outcomes. The OS at 5 years of the mLVRH is superior to the TLRH. The DFS at 5 years in Ⅰb1 stage, the mLVRH is higher than the TLRH. Therefore, the modified laparoscopy is still an alternative surgery for early cervical cancer patients when following the principle of no-tumor-exposure.
9.The expression and the diagnostic value of NOD2 in hepatocellular carcinoma
Jingxin ZHANG ; Lili QIAO ; Ning LIANG ; Jian XIE ; Hui LUO ; Guodong DENG ; Jiandong ZHANG
Journal of International Oncology 2016;43(7):499-502
Objective To investigate the expression of nucleotide-binding oligomerization domain protein 2 (NOD2)in serum of patients with hepatocellular carcinoma (HCC),and to analyse the roles of NOD2 in HCC development and its clinical diagnostic value.Methods This study including 66 patients with HCC in the hospi-tal from March 1,2013 to December 31,2014 and 61 healthy controls.Serum NOD2 levels were determined by enzyme-linked immunosorbent assay (ELISA).Analysis of significance was performed with rank sum test using SPSS statistical 16.0 software.Results Serum levels of NOD2 in HCC patients were 171 pg/ml,significantly higher than that of healthy controls(95 pg/ml,Z =-5.00,P =0.00),and the serum NOD2 levels were correla-ted with clinical stage of HCC (H=56.26,P =0.00).Compared with the serum NOD2 levels in stageⅠ,Ⅱpatients (106 pg/ml)and healthy controls (95 pg/ml),the serum NOD2 level in stage Ⅲ and Ⅳ (220 pg/ml) were significantly increased (χ2 =31.24,P =0.00;χ2 =47.23,P =0.00),but the expression of NOD2 in stageⅠandⅡwere nearly equal to that of the healthy controls (χ2 =0.36,P =0.83).The ROC analysis revealed that the best diagnostic cutoff-point of serum NOD2 levels for predicting the Ⅲ and Ⅳ stages of HCC was 148.78 pg/ml,meanwhile corresponding sensitivity was 89.1% and specificity was 77.0%.Additionally,corre-lation analysis demonstrated that there was no significant correlation between NOD2 and alpha-fetal protein (r =0.44,P =0.14).Survival curves obtained that the survival time of HCC patients with NOD2 serum concentrations≥ 200 pg/ml was significantly less than that <200 pg/ml (χ2 =15.32,P <0.05).Conclusion NOD2 is highly expressed in the serum of HCC patients,especially in advanced patients,which is possibly involved in the development of HCC and has the potential to become an effective marker used for HCC diagnosis.

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