1.High Expression of RPL22L1 in Colorectal Cancer and its Correlation With Patients' Poor Prognosis and Glucose Metabolism of Tumor Cells
Shasha CAI ; Changfa YU ; Yaping JIANG ; Darong DUAN ; Han FANG ; Wenxiao CHEN ; Jinxing XIA
Chinese Journal of Gastroenterology 2025;30(3):139-145
Background:Ribosomal protein L22-like 1(RPL22L1)exerts regulatory effects on various malignant tumors such as lung cancer,prostate cancer,and cervical cancer.However,its role in colorectal cancer(CRC)remains unclear.Aims:To investigate the expression of RPL22L1 in CRC and its role in patients' prognosis and glucose metabolism of tumor cells.Methods:A total of 142 newly diagnosed CRC patients admitted to the Taizhou First People's Hospital from February 2022 to June 2024 were enrolled.The expression levels of RPL22L1 mRNA and protein were detected by quantitative real-time PCR and immunohistochemistry,respectively.The correlation between RPL22L1 expression and clinicopathological characteristics was analyzed.Kaplan-Meier survival analysis was used to evaluate the impact of RPL22L1 expression on the prognosis of CRC patients.RPL22L1 siRNA was transfected into SW480 cells to establish a low-expression cell model.Cell proliferation was assessed by CCK-8 assay,cell migration by Transwell chamber assay,and apoptosis by flow cytometry.Gene set enrichment analysis(GSEA)was performed to evaluate the effect of RPL22L1 on glucose metabolism of tumor cells.Results:The expression levels of RPL22L1 mRNA and protein were significantly higher in CRC tissues than in adjacent normal tissues(all P<0.05).The expression level of RPL22L1 mRNA was correlated with the TNM stage and carcinoembryonic antigen level of CRC(all P<0.05).Kaplan-Meier analysis showed that the cumulative survival rate of high RPL22L1 mRNA expression group was significantly lower than that of low-expression group(P=0.027).The expression level of RPL22L1 mRNA was significantly higher in SW480 cells than in normal intestinal epithelial cells(P<0.001).After inhibiting RPL22L1 expression,the proliferation and migration capacities of SW480 cells were significantly decreased(all P<0.05),the apoptosis rate was significantly increased(P=0.005),and the lactate level and relative glucose uptake level were significantly reduced(all P<0.05).GSEA indicated that RPL22L1 gene was associated with glycolysis/gluconeo-genesis(P=0.02).Conclusions:RPL22L1 is highly expressed in CRC and is associated with poor prognosis of patients,suggesting its potential as a molecular target for CRC therapy.Furthermore,RPL22L1 may promote the tumorigenesis and progression of CRC by modulating glucose metabolism.
2.High Expression of RPL22L1 in Colorectal Cancer and its Correlation With Patients' Poor Prognosis and Glucose Metabolism of Tumor Cells
Shasha CAI ; Changfa YU ; Yaping JIANG ; Darong DUAN ; Han FANG ; Wenxiao CHEN ; Jinxing XIA
Chinese Journal of Gastroenterology 2025;30(3):139-145
Background:Ribosomal protein L22-like 1(RPL22L1)exerts regulatory effects on various malignant tumors such as lung cancer,prostate cancer,and cervical cancer.However,its role in colorectal cancer(CRC)remains unclear.Aims:To investigate the expression of RPL22L1 in CRC and its role in patients' prognosis and glucose metabolism of tumor cells.Methods:A total of 142 newly diagnosed CRC patients admitted to the Taizhou First People's Hospital from February 2022 to June 2024 were enrolled.The expression levels of RPL22L1 mRNA and protein were detected by quantitative real-time PCR and immunohistochemistry,respectively.The correlation between RPL22L1 expression and clinicopathological characteristics was analyzed.Kaplan-Meier survival analysis was used to evaluate the impact of RPL22L1 expression on the prognosis of CRC patients.RPL22L1 siRNA was transfected into SW480 cells to establish a low-expression cell model.Cell proliferation was assessed by CCK-8 assay,cell migration by Transwell chamber assay,and apoptosis by flow cytometry.Gene set enrichment analysis(GSEA)was performed to evaluate the effect of RPL22L1 on glucose metabolism of tumor cells.Results:The expression levels of RPL22L1 mRNA and protein were significantly higher in CRC tissues than in adjacent normal tissues(all P<0.05).The expression level of RPL22L1 mRNA was correlated with the TNM stage and carcinoembryonic antigen level of CRC(all P<0.05).Kaplan-Meier analysis showed that the cumulative survival rate of high RPL22L1 mRNA expression group was significantly lower than that of low-expression group(P=0.027).The expression level of RPL22L1 mRNA was significantly higher in SW480 cells than in normal intestinal epithelial cells(P<0.001).After inhibiting RPL22L1 expression,the proliferation and migration capacities of SW480 cells were significantly decreased(all P<0.05),the apoptosis rate was significantly increased(P=0.005),and the lactate level and relative glucose uptake level were significantly reduced(all P<0.05).GSEA indicated that RPL22L1 gene was associated with glycolysis/gluconeo-genesis(P=0.02).Conclusions:RPL22L1 is highly expressed in CRC and is associated with poor prognosis of patients,suggesting its potential as a molecular target for CRC therapy.Furthermore,RPL22L1 may promote the tumorigenesis and progression of CRC by modulating glucose metabolism.
3.Nodosin Induces Apoptosis of Colorectal Cancer Cells by Inhibiting mTOR Pathway and Activating Autophagy
Hong ZHANG ; Shasha CAI ; Chunqin XIE ; Changfa YU ; Han FANG
Chinese Journal of Gastroenterology 2024;29(5):291-296
Background:Colorectal cancer is a common malignant tumor in the digestive tract.The existing treatment methods and drugs have disadvantages such as significant toxicity and side effects,so there is an urgent need to develop new drugs with high efficiency and low toxicity.Aims:To investigate the potential mechanism of Nodosin in inducing apoptosis of colorectal cancer cells.Methods:Colorectal cancer cell lines SW480 and HCT116 were cultured with 0,5,10 and 20 μmol/L Nodosin for 24 hours,or cultured with 10 μmol/L Nodosin for 0,12,24 and 48 hours.The cell viability was measured by CCK-8 assay,and the protein expressions of PARP1,mTOR,p-mTOR and LC3BⅡ were detected by Western blotting.After pretreated with the autophagy inhibitor 3-methyladenine(3-MA)5 mmol/L for 4 hours,SW480 and HCT116 cells were cultured with 10 μmol/L Nodosin for 24 hours.The protein expressions of PARP1 and LC3BⅡ were detected by Western blotting.Results:Nodosin could decrease the cell viability of SW480 and HCT116 cells in a dose-and time-dependent manner,increase the expressions of apoptosis-related protein cleaved PARP1 and autophagy-related protein LC3BⅡ,and inhibit the protein expression of p-mTOR.Pretreatment of the autophagy inhibitor 3-MA could inhibit the expressions of apoptosis-related protein cleaved PARP1 and autophagy-related protein LC3BⅡ induced by Nodosin.Conclusions:Nodosin can induce apoptosis of colorectal cancer cells by inhibiting mTOR pathway and activating autophagy.
4.Progress in the optimization of CRISPR/Cas9 technology for the establishment of genetically modified mouse models
Yu WANG ; Yong WU ; Chunnan LIANG ; Changfa FAN
Chinese Journal of Comparative Medicine 2024;34(5):152-158
CRISPR/Cas9 technology has driven the development of various fields in life science.With continuous deepening of its understanding,researchers have made multiple improvements and optimizations to adapt to various application scenarios.The optimization of CRISPR/Cas9 technology has also provided breakthroughs in the establishment of genetically modified mouse models.This article briefly reviews the development process of CRISPR/Cas9 technology and summarizes optimization strategies of CRISPR/Cas9,establishment of conditional knockout/knockin gene-modified mouse models,and delivery systems for CRISPR/Cas9 elements and HDR templates.
5.GANT61 Induces Colon Cancer Cell Apoptosis by Regulating Autophagy
Hong ZHANG ; Jianguo ZHANG ; Miao HE ; Zheng YUAN ; Han FANG ; Changfa YU ; Qingqing XIA
Chinese Journal of Gastroenterology 2024;29(12):731-736
Background:Colorectal cancer is a common malignant tumor of the digestive system,its incidence rate is increasing and seriously affect human health.Aims:To investigate whether GANT61,an inhibitor of Hedgehog signaling pathway,induces apoptosis of colon cancer cells by regulating autophagy.Methods:Colon cancer cell line SW480 was cultured in vitro and treated with GANT61 of different concentrations(0,10,20,and 40 μmol/L)for 24 h.The cell viability was measured by CCK-8 assay,and the apoptosis rate was determined by flow cytometry.Expressions of the key molecules in Hedgehog/GLI signaling pathway,as well as the proteins related to autophagy and apoptosis were detected by Western blotting and/or qRT-PCR.Then the SW480 cells were treated with combination of GANT61 and chloroquine,an autophagy inhibitor,or rapamycin,an autophagy agonist,the alterations of the apoptosis rates and the related protein expressions were assessed.Results:The viability of SW480 cells decreased and the apoptosis rate increase with the increase of GANT61 concentration.Meanwhile,the mRNA and protein expressions of GLI1,GLI2,cyclin D1(a cell cycle G1/S-specific protein),and BCL-2(an anti-apoptotic protein)were inhibited,and the protein expressions of C-PARP1(an apoptosis-related protein)and BECLIN1,LC3Ⅱ(autophagy-related proteins)were increased.Chloroquine could partially reverse the effect of GANT61 on autophagy activation and cell apoptosis,while rapamycin could promote the pro-apoptotic effect of GANT61.Conclusions:GANT61 can activate autophagy and induce apoptosis in colon cancer cells.
6.Nodosin Induces Apoptosis of Colorectal Cancer Cells by Inhibiting mTOR Pathway and Activating Autophagy
Hong ZHANG ; Shasha CAI ; Chunqin XIE ; Changfa YU ; Han FANG
Chinese Journal of Gastroenterology 2024;29(5):291-296
Background:Colorectal cancer is a common malignant tumor in the digestive tract.The existing treatment methods and drugs have disadvantages such as significant toxicity and side effects,so there is an urgent need to develop new drugs with high efficiency and low toxicity.Aims:To investigate the potential mechanism of Nodosin in inducing apoptosis of colorectal cancer cells.Methods:Colorectal cancer cell lines SW480 and HCT116 were cultured with 0,5,10 and 20 μmol/L Nodosin for 24 hours,or cultured with 10 μmol/L Nodosin for 0,12,24 and 48 hours.The cell viability was measured by CCK-8 assay,and the protein expressions of PARP1,mTOR,p-mTOR and LC3BⅡ were detected by Western blotting.After pretreated with the autophagy inhibitor 3-methyladenine(3-MA)5 mmol/L for 4 hours,SW480 and HCT116 cells were cultured with 10 μmol/L Nodosin for 24 hours.The protein expressions of PARP1 and LC3BⅡ were detected by Western blotting.Results:Nodosin could decrease the cell viability of SW480 and HCT116 cells in a dose-and time-dependent manner,increase the expressions of apoptosis-related protein cleaved PARP1 and autophagy-related protein LC3BⅡ,and inhibit the protein expression of p-mTOR.Pretreatment of the autophagy inhibitor 3-MA could inhibit the expressions of apoptosis-related protein cleaved PARP1 and autophagy-related protein LC3BⅡ induced by Nodosin.Conclusions:Nodosin can induce apoptosis of colorectal cancer cells by inhibiting mTOR pathway and activating autophagy.
7.GANT61 Induces Colon Cancer Cell Apoptosis by Regulating Autophagy
Hong ZHANG ; Jianguo ZHANG ; Miao HE ; Zheng YUAN ; Han FANG ; Changfa YU ; Qingqing XIA
Chinese Journal of Gastroenterology 2024;29(12):731-736
Background:Colorectal cancer is a common malignant tumor of the digestive system,its incidence rate is increasing and seriously affect human health.Aims:To investigate whether GANT61,an inhibitor of Hedgehog signaling pathway,induces apoptosis of colon cancer cells by regulating autophagy.Methods:Colon cancer cell line SW480 was cultured in vitro and treated with GANT61 of different concentrations(0,10,20,and 40 μmol/L)for 24 h.The cell viability was measured by CCK-8 assay,and the apoptosis rate was determined by flow cytometry.Expressions of the key molecules in Hedgehog/GLI signaling pathway,as well as the proteins related to autophagy and apoptosis were detected by Western blotting and/or qRT-PCR.Then the SW480 cells were treated with combination of GANT61 and chloroquine,an autophagy inhibitor,or rapamycin,an autophagy agonist,the alterations of the apoptosis rates and the related protein expressions were assessed.Results:The viability of SW480 cells decreased and the apoptosis rate increase with the increase of GANT61 concentration.Meanwhile,the mRNA and protein expressions of GLI1,GLI2,cyclin D1(a cell cycle G1/S-specific protein),and BCL-2(an anti-apoptotic protein)were inhibited,and the protein expressions of C-PARP1(an apoptosis-related protein)and BECLIN1,LC3Ⅱ(autophagy-related proteins)were increased.Chloroquine could partially reverse the effect of GANT61 on autophagy activation and cell apoptosis,while rapamycin could promote the pro-apoptotic effect of GANT61.Conclusions:GANT61 can activate autophagy and induce apoptosis in colon cancer cells.
8.Clinical features of the predilection and severer sites of intrauterine adhesions.
Yang YU ; Lingxiao ZOU ; Waixing LI ; Xingping ZHAO ; Changfa SHU ; Chunxia CHENG ; Dabao XU
Journal of Central South University(Medical Sciences) 2022;47(11):1568-1574
OBJECTIVES:
Intrauterine adhesions (IUA) refers to the adhesions between the myometrium of the uterine cavity, which is secondary to damage to the basal layer of the endometrium due to trauma or infection. The occurrence of IUA is mainly related to intrauterine operations. Hysteroscopic adhesiolysis (HA) is the standard surgical treatment for IUA. But the recurrence rate of IUA after HA is still high. Importantly, endometrium recovery is difficult, resulting in unsatisfied prognosis for moderate to severer IUA patients. Therefore, it is important to take effective primary preventive measures against the etiology to avoid endometrium damage from medical surgery. In this paper, we discuss and analyze predilection and severer sites of intrauterine adhesions, aiming to provide a basis for how to avoid and reduce injuries during intrauterine operations, such as abortion, dilation and curettage.
METHODS:
In this study, we retrospectively analyzed the surgical videos of patients who underwent HA for the first time from January 2019 to December 2021 in the Third Xiangya Hospital of Central South University so as to assess the area of adhesions and predilection and severer sites of occurrence of adhesions, and we collected 657 patients who underwent HA for the first time, including 81 patients with total IUA and 576 patients with partial IUA. We counted and analyzed the number and composition ratio of partial IUA patients with severer sites of damage to the lateral wall of the uterine cavity and severerr sites of damage to each segment of the uterine cavity.
RESULTS:
Among 576 patients with partial IUA, there were 60 patients with no significant difference in the degree of adhesions between the right and left sides, 143 patients with severer adhesions on the left side of the uterine cavity, and 373 patients with severer adhesions on the right side of the uterine cavity. There was a difference in the severity of damage of left and right lateral wall. The proportion of patients with severer adhesions on the right side of the uterine cavity (64.8%) was higher than that of patients with adhesions on the left side of the uterine cavity (24.8%), and there was statistically difference (P<0.05). There was 93 patients with severer adhesions at the fundus or bilateral horn of the uterus, 190 patients with severer adhesions at the middle and upper part of the uterine cavity, 245 patients with severer adhesions at the middle and lower part of the uterine cavity and at the endocervix, and 48 patients with no significant difference in the degree of adhesions in each part. The proportion of patients with severer adhesions at the middle and lower part of the uterine cavity and at the endocervix was higher (42.5%) than those with adhesions in the fundus or bilateral horn of the uterus (16.1%) and in the middle and upper part of the uterine cavity (33.0%), and there were statistically differences (both P<0.05).
CONCLUSIONS
The predilection site of IUA is the lateral wall of the uterine cavity. The severer adhesions is in the right lateral wall of the uterine cavity, the middle and lower segments and the endocervix, which may be related to the operating habits of the surgeon. Therefore, gynecologists should minimize damage to the lateral wall of the uterine cavity, especially the right lateral wall in performing uterine operations (more attention should be paid by right-handed physicians). Besides, we should pay attention to protecting the middle and lower segments of the uterine cavity and the endocervix, avoiding maintaining negative pressure to withdraw the uterine tissue suction tube from the uterine cavity during abortion procedures to minimize damage.
Humans
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Retrospective Studies
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Tissue Adhesions
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Uterus/pathology*
9.Diagnostic Value of Detection of 16S rRNA Gene of Pathogens from Blood by PCR
Changfa YU ; Lijun YE ; Yingpeng REN ; Darong DUAN ; Ronghua RUAN ; Xiansen ZHANG
Chinese Journal of Nosocomiology 2009;0(19):-
OBJECTIVE To explore study a method for rapid detection of bacterial infection in clinic to diagnose septicemia early.METHODS 16S rRNA gene of ten bacterial species was amplified with PCR,by using human genome DNA,HBV-DNA and Candida albicans as comparison.The sensitivity test was done by the method of gradual dilution of Escherichia coli.RESULTS The bacterial species were amplified and the products were 371 bp,but human genome DNA,HBV-DNA and C.albicans showed no amplification products.Sensitivity test showed that it could detect as low as 1.5?104/L of E.coli.CONCLUSIONS The method is rapid and highly specific and sensitive in detecting the existence of bacterial 16S rRNA gene.
10.Application of Polymerase Chain Reaction in Detection of Pathogens in Cerebrospinal Fluid
Xiansen ZHANG ; Changfa YU ; Darong DUAN ; Yingpeng REN ; Ronghua RUAN ; Xiaoming YING
Chinese Journal of Nosocomiology 2009;0(22):-
OBJECTIVE To set up a quick method to detect pathogens in cerebrospinal fluid.METHODS The method of polymerase chain reaction(PCR),using a pair of universal primers targeted at the 16S rRNA gene,was adopted to amplify the DNA of bacterium.138 clinical specimens obtained from patients were examined by PCR method and bacterial culture method.RESULTS The positive rate was 39.86% of PCR and 17.39% of culture,the outcomes had statistical significance(P

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