1.Combination of brefeldin A and tunicamycin induces apoptosis in HepG2 cells through the endoplasmic reticulum stress-activated PERK-eIF2α-ATF4-CHOP signaling pathway
Minghong LI ; Mengyi DUAN ; Ying YANG ; Xingdao LI ; Dan LI ; Wenting GAO ; Xiaotong JI ; Jianying BAI
Liver Research 2025;9(1):49-56
Background and aims:Hepatocellular carcinoma(HCC)is a malignant tumor with a high mortality rate,but there are still no effective treatments.The aim of this study was to investigate the anticancer po-tential of the combined use of brefeldin A(BFA)and tunicamycin(TM)in HepG2 cells,as well as the underlying mechanisms.Methods:HepG2 cells were treated with different concentrations of BFA(0.1-2.5 mg/L)and TM(1-5 mg/L)for 24 h.DMSO(0.1%,v/v)was used as a vehicle control.Cell viability and cell migration were measured using MTT assay and scratch wound assay,respectively.Apoptosis was detected using flow cytometry and acridine orange(AO)staining.The protein and mRNA levels of various factors involved in apoptosis(poly(ADP-ribose)polymerase-1(PARP-1),caspase-12,caspase-3,and stearoyl-CoA desaturase 1)and endoplasmic reticulum(ER)stress(binding immunoglobulin protein(BiP),protein kinase R-like endoplasmic reticulum kinase(PERK),p-PERK,phosphorylation of eukaryotic translation initiation factor 2alpha(p-eIF2α),activating transcription factor(ATF)4,and C/EBP homologous protein(CHOP))were measured using Western blotting and qRT-PCR,respectively.Results:Both BFA and TM alone significantly reduced the viability of HepG2 cells in a dose-dependent way.The co-incubation with TM(1 mg/L)further significantly reduced the viability of HepG2 cells treated with BFA(0.25 mg/L)alone(P<0.05).BFA significantly increased the protein and mRNA levels of caspase-3 and PARP-1(P<0.05)compared to control and DMSO-treated cells,indicating that BFA induced apoptosis in HepG2 cells by increasing the expression of caspase-3 and PARP-1.The induction of apoptosis by BFA could be further significantly enhanced by co-incubation with TM.In addition,BFA significantly increased the mRNA levels of BiP,PERK and ATF4(P<0.05)compared to control and DMSO-treated cells.After co-incubation of BFA and TM,the protein levels of BiP,p-PERK,p-eIF2α and CHOP were significantly increased,indicating that TM could enhance BFA-induced ER stress in HepG2 cells through the PERK-eIF2α-ATF4-CHOP pathway.Conclusions:BFA could induce apoptosis and ER stress,and TM could enhance the ability of BFA to induce apoptosis and ER stress in HepG2 cells through the PERK-eIF2α-ATF4-CHOP pathway.The findings highlight the therapeutic potential of the combined use of BFA and TM in treating HCC.
2.Cross-cultural adaptation and validation of the postoperative recovery in children scale for dental treatment under general anesthesia (PRiC-DTGA) in Chinese children
Xiaotong SHI ; Bing LIU ; Ze FAN ; Yao ZHANG ; Tiantian LIU ; Zhenzhen GAO ; Haopeng ZHANG ; Hui ZHANG
Chinese Journal of Stomatology 2025;60(10):1128-1136
Objective:To translate and adapt the postoperative recovery in children (PRiC) scale, developing a Chinese version for children undergoing dental treatment under general anesthesia (PRiC-DTGA) with validated psychometric properties.Methods:The PRiC scale underwent forward-backward translation using Brislin′s model. A convenience sample of DTGA patients from the Department of Anesthesiology, School of Stomatology, The Fourth Mility Force Medical University was enrolled for a cross-sectional survey on postoperative complications. Delphi expert consultation informed cultural adaptation based on survey findings to develop the PRiC-DTGA Chinese version. Psychometric validation included reliability and validity testing in a separate DTGA cohort at the same center (April-October 2024).Results:Results from the cross-sectionalsurvey of 231 children showed that 82.7% (191/231) of them hadat least one postoperative complication within 72 hours, and these complications were mainly mild local symptoms. Additionally, 358 copies of the Chinese version of the PRiC-DTGA scale were distributed; 21 invalid questionnaires with incomplete information were excluded, and a total of 337 cases were included inthe study. The final PRiC-DTGA comprised 22 items across three dimensions including physical comfort, social ability, and negative emotional. Exploratory factor analysis (EFA) confirmed all factor loadings>0.4. Confirmatory factor analysis (CFA) demonstrated adequate fit: χ 2/df=1.665, tucker-Lewis index (TLI)=0.924, comparative fit index (CFI)=0.896, standardized root mean square residual (SRMR)=0.041, and root mean square error of approximation (RMSEA)=0.044 (90% CI: 0.035-0.053). Reliability was strong with Cronbach′s α (total scale)=0.853, subscale α=0.632-0.723, split-half reliability=0.824. Validity indices met standards: scale-content validity index (S-CVI)=0.909, Item-CVI range=0.944-1.000, average variance extracted (AVE)=0.473-0.501, composite reliability (CR)=0.830-0.913. Conclusions:The systematically adapted PRiC-DTGA demonstrates robust reliability and validity, serving as an effective tool for assessing postoperative recovery quality in Chinese children following DTGA.
3.Immune-metabolic regulatory mechanism of the gut-eye axis of intestinal flora in dry eye and new treatment strategies
Xiaotong LIU ; Xinyi SUN ; Weiping GAO
International Eye Science 2025;25(11):1771-1776
Dry eye disease is a multifactorial chronic ocular surface disorder that is challenging to manage because its pathogenesis is not well understood. The gut-eye axis theory provides insights into its pathogenesis to guide prevention and treatment. Intestinal flora dysbiosis induces dry eye disease through complex mechanisms involving inflammation, ocular surface microbiota, nerve damage, and microbial metabolites. This article reviews the immunometabolic regulation of the gut-eye axis and summarizes gut flora-targeted interventions(fecal microbiota transplantation, probiotic supplementation, and dietary modification)to provide a theoretical basis for dry eye disease prevention and treatment.
4.Effect of LINC01355 on the proliferation,apoptosis,and invasion of oral squamous cell carcinoma cells through miR-545-5p/FOXD1 signaling pathway
Jing GAO ; Xiaotong WEI ; Xingchen LI ; Wei YAN ; Hao WANG
Journal of China Medical University 2025;54(3):238-245
Objective To investigate the effects of LINC01355 on the proliferation,apoptosis,and invasion of oral squamous cell car-cinoma(OSCC)cells via the miR-545-5p/forkhead box D1(FOXD1)signaling pathway.Methods Cal-27 cells were cultured in vitro and randomly separated into the control group,si-LINC01355(transfected with LINC01355 siRNA)group,pc-LINC01355(transfected with LINC01355 overexpression plasmid)group,si-NC+miR-545-5p-NC+pc-NC(transfected with LINC01355 siRNA negative control,miR-545-5p negative control and empty plasmid)group,and si-LINC01355+miR-545-5p inhibitor(transfected with LINC01355 siRNA and miR-545-5p inhibitor)group.After transfection,quantitative real-time PCR was applied to detect the expression of LINC01355,miR-545-5p,and FOXD1 in cells.The Cal-27 cells transfected into groups were then subcutaneously inoculated to construct nude mouse models of transplanted tumors in each group,and the growth of the transplanted tumors was measured.The CCK-8 method,flow cytom-etry,and Transwell assay were applied in order to detect cell proliferation,apoptosis,and invasion,respectively.Immunohistochemical staining was applied to detect the expression of epithelial-mesenchymal transition(EMT)related proteins Vimentin and E-cadherin in cells.Western blotting was applied to detect the expression of cell proliferation related proteins(proliferating cell nuclear antigen[PCNA],C-myc),apoptosis related proteins(cleaved cysteinyl aspartate-specific proteases-3[cleaved caspase-3],BCL-2-associated X protein[Bax]),and FOXD1 protein.A dual-luciferase reporter assay was used to identify the relationship between LINC01355 and the miR-545-5p/FOXD1 signaling pathway.Results Compared with the control group,the expression of LINC01355 and FOXD1 mRN A,proliferation activity,number of invasions,positive expression of Vimentin protein,expression of PCNA,C-myc and FOXD1 protein,and transplanted tumor volume reduced(All P<0.05);the expression of miR-545-5p,apoptosis rate,cleaved caspase-3 and Bax protein expression,and positive expression of E-cadherin protein increased(All P<0.05)in the si-LINC01355 group.The trend of changes in the various indica-tors of the pc-LINC01355 group was opposite to that of the si-LINC01355 group.Compared with the si-LINC01355 group,the expression of LINC01355 and FOXD1 mRNA,prolife ration activity,number of invasions,positive expression of Vimentin protein,expression of PCNA,C-myc and FOXD1 proteins,and the transplanted tumor volume in the si-LINC01355+miR-545-5p inhibitor group increased(All P<0.05),whereas the expression of miR-545-5p,apoptosis rate,cleaved caspase-3 and Bax protein expression,and positive expression of E-cadherin protein decreased(All P<0.05).LINC01355 was able to downregulate miR-545-5p expression in Cal-27 cells and miR-545-5p was able to downregulate FOXD1 expression.Conclusion LINC01355 promotes OSCC cell proliferation and invasion,and inhibits apoptosis by targeting the miR-545-5p/FOXD1 signaling pathway.
5.The relationship between SERPINA4 expression and prognosis in metastatic color-ectal cancer
Jianfang GAO ; Xiaotong DONG ; Qing ZHAO ; Yunxiang ZHANG ; Huimin SUN
Chinese Journal of Clinical and Experimental Pathology 2025;41(3):340-344,351
Purpose To investigate the relationship between SERPINA4 expression and clinicopathological fea-tures and prognosis in metastatic colorectal cancer(CRC).Methods EnVision two-step immunohistochemical method was used to detect the expression of SERPINA4 protein in 104 cases of primary and metastatic tumor paraffin tissues of metastatic colorectal cancer and adjacent intestinal normal mucosa,and its relationship with clinicopathological features of patients was analyzed.qRT-PCR was used to detect the expression of SERPINA4 mRNA in 11 pairs of metastatic CRC metastases and paired primary tumor paraffin-embedded tissue samples.The survival of patients was followed up,and the relationship between SERPINA4 expression and patient prognosis was analyzed by Kaplan-Meier method.Re-sults Immunohistochemical results showed that the strong positive expression rate of SERPINA4 was 97.10%in nor-mal mucosa,71.20%in primary lesions,and 39.40%in metastatic lesions.The difference between the three groups was statistically significant(P<0.001).qRT-PCR results showed that SERPINA4 mRNA was lowly expressed in me-tastatic lesions of metastatic CRC(P<0.05).The expression of SERPINA4 in metastatic lesions was significantly cor-related with histological grade,N stage and M stage of the tumor(P<0.05),but not significantly correlated with age,gender,tumor location and T stage of patients(P>0.05).The survival time of patients with metastatic colorectal cancer with high expression of SERPINA4 was longer than that of patients with low expression.Conclusion The ex-pression of SERPINA4 is of great significance in the survival and prognosis of patients with metastatic CRC.
6.Cross-cultural adaptation and validation of the postoperative recovery in children scale for dental treatment under general anesthesia (PRiC-DTGA) in Chinese children
Xiaotong SHI ; Bing LIU ; Ze FAN ; Yao ZHANG ; Tiantian LIU ; Zhenzhen GAO ; Haopeng ZHANG ; Hui ZHANG
Chinese Journal of Stomatology 2025;60(10):1128-1136
Objective:To translate and adapt the postoperative recovery in children (PRiC) scale, developing a Chinese version for children undergoing dental treatment under general anesthesia (PRiC-DTGA) with validated psychometric properties.Methods:The PRiC scale underwent forward-backward translation using Brislin′s model. A convenience sample of DTGA patients from the Department of Anesthesiology, School of Stomatology, The Fourth Mility Force Medical University was enrolled for a cross-sectional survey on postoperative complications. Delphi expert consultation informed cultural adaptation based on survey findings to develop the PRiC-DTGA Chinese version. Psychometric validation included reliability and validity testing in a separate DTGA cohort at the same center (April-October 2024).Results:Results from the cross-sectionalsurvey of 231 children showed that 82.7% (191/231) of them hadat least one postoperative complication within 72 hours, and these complications were mainly mild local symptoms. Additionally, 358 copies of the Chinese version of the PRiC-DTGA scale were distributed; 21 invalid questionnaires with incomplete information were excluded, and a total of 337 cases were included inthe study. The final PRiC-DTGA comprised 22 items across three dimensions including physical comfort, social ability, and negative emotional. Exploratory factor analysis (EFA) confirmed all factor loadings>0.4. Confirmatory factor analysis (CFA) demonstrated adequate fit: χ 2/df=1.665, tucker-Lewis index (TLI)=0.924, comparative fit index (CFI)=0.896, standardized root mean square residual (SRMR)=0.041, and root mean square error of approximation (RMSEA)=0.044 (90% CI: 0.035-0.053). Reliability was strong with Cronbach′s α (total scale)=0.853, subscale α=0.632-0.723, split-half reliability=0.824. Validity indices met standards: scale-content validity index (S-CVI)=0.909, Item-CVI range=0.944-1.000, average variance extracted (AVE)=0.473-0.501, composite reliability (CR)=0.830-0.913. Conclusions:The systematically adapted PRiC-DTGA demonstrates robust reliability and validity, serving as an effective tool for assessing postoperative recovery quality in Chinese children following DTGA.
7.Three-dimensional radiographic characteristics of 18 odontogenic myxoma
Liuyang QU ; Xiaotong LING ; Danni ZHENG ; Jing YANG ; Xuebing YAN ; Denggao LIU ; Yan GAO
Journal of Practical Stomatology 2025;41(5):680-684
Objective:To analyze the there-dimensional radiographic characteristics of odontogenic myxoma(OM).Methods:Clinical records,histopathological reports,SCT or CBCT images of 18 patients with OM were retrospectively collected,and radiograph-ic features including lesion site,shape,size,expansion,intralesional septa and relationship with surrounding tissues were analyzed.The lesions were classified into types by the forms of septa.Results:18 patients(8 males and 10 females,aged 9-39 years old)were included.Among them,12 lesions(66.7%)were round-like in shape,7(38.9%)were located in the posterior maxilla,and 8(44.4%)in the posterior mandible.According to the septa map,the lesions included 4(22.2%)without septum(unilocular type),4 with(22.2%)sporadic septum on the periphery of the tumor(peripheral type),4(22.2%)with the septum from the periphery towards the center of the tumor(centripetal type)and 6(16.7%)with multiple septa interwoven as grid-like trabeclations(criss-crossing tup).Labial/buccal cortex expansion was present in 11 lesions(61.1%),and palatal cortex expansion in 12 lesions(66.7%).Moreover,cortex discontinuity occurred in 14 lesions(77.8%),impacted tooth was present in 9 lesions(16.7%),and root resorption was present in 8 lesions(44.4%).Conclusion:Most OMs occur in the posterior jaw,with multilocular and discontinued cortex.OMs can be classified into unilocular,perpheral,centripetal and cirss-crossing type by the forms of septa in the lesion.
8.Evaluation of the efficacy of dienogest in the treatment of adenomyosis with different MRI types
Molin WANG ; Hongyan GUO ; Xinran GAO ; Lu LIU ; Xiaotong HAN
Chinese Journal of Obstetrics and Gynecology 2025;60(7):511-519
Objective:To analyze the differences in the clinical characteristics of patients with adenomyosis of different magnetic resonance imaging (MRI) types and the differences in treatment effects after the application of dienogest.Methods:A total of 176 patients with adenomyosis who were admitted to Peking University Third Hospital from June 2017 to February 2023 were included in the study, and all of them were clearly classified by pelvic MRI and treated with dienogest. The clinical characteristics and treatment of the patients were retrospectively collected, and the patients were divided into endogenous type, exogenous type and penetrating type by MRI. The differences in clinical symptoms, imaging features and treatment effect of patients with adenomyosis of different MRI types were analyzed.Results:(1) The percentages of patients with endogenous, exogenous, and penetrating types were 40.9% (72/176), 35.2% (62/176) and 23.9% (42/176), respectively. The proportion of dysmenorrhea in patients with endogenous type (90.3%, 65/72) was significantly lower than those of exogenous type (100.0%, 62/62) and penetrating type (97.6%, 41/42; χ2=7.853, P=0.020), while there was no significant difference between exogenous type and penetrating type ( P>0.05). There were no statistically significant differences in menarche time, menstrual cycle and menstrual period among the three types of patients (all P>0.05), there was also no statistically significant difference in the proportion of menstrual abnormalities (including heavy and irregular menstrual bleeding; P>0.05). The proportions of ovarian endometrioma and deep infiltrating endometriosis in exogenous and penetrating types were significantly higher than that in endogenous type (all P<0.05). (2) The pain scores of all patients were significantly lower than those before treatment (all P<0.001), the proportion of patients with exogenous type (62.9%, 39/62) who had complete remission after treatment was higher than those of endogenous type (49.2%, 32/65) and penetrating type (46.3%, 19/41), but there was no significant difference in pain relief (i.e. the variation in the pain scores) between the three types ( P>0.05). (3) Endogenous type ( OR=0.361, 95% CI: 0.147-0.883; P=0.026), failure to apply gonadotropin-releasing hormone agonist (GnRH-a) in advance ( OR=0.208, 95% CI: 0.083-0.518; P<0.001), cystic changes ( OR=2.671, 95% CI: 1.108-6.437; P=0.029) and abnormal menstruation ( OR=3.466, 95% CI: 1.464-8.209; P=0.005) were independent risk factors for irregular bleeding after dienogest treatment. Conclusions:(1) There are obvious differences in the clinical characteristics of patients with adenomyosis of different MRI types, and patients with exogenous and penetrating types are more likely to have dysmenorrhea symptoms. (2) Dienogest could significantly alleviate the symptoms of dysmenorrhea in patients with adenomyosis. (3) Endogenous type, failure to take GnRH-a in advance and associated menstrual abnormalities are independent risk factors for irregular bleeding after dienogest treatment.
9.The relationship between SERPINA4 expression and prognosis in metastatic color-ectal cancer
Jianfang GAO ; Xiaotong DONG ; Qing ZHAO ; Yunxiang ZHANG ; Huimin SUN
Chinese Journal of Clinical and Experimental Pathology 2025;41(3):340-344,351
Purpose To investigate the relationship between SERPINA4 expression and clinicopathological fea-tures and prognosis in metastatic colorectal cancer(CRC).Methods EnVision two-step immunohistochemical method was used to detect the expression of SERPINA4 protein in 104 cases of primary and metastatic tumor paraffin tissues of metastatic colorectal cancer and adjacent intestinal normal mucosa,and its relationship with clinicopathological features of patients was analyzed.qRT-PCR was used to detect the expression of SERPINA4 mRNA in 11 pairs of metastatic CRC metastases and paired primary tumor paraffin-embedded tissue samples.The survival of patients was followed up,and the relationship between SERPINA4 expression and patient prognosis was analyzed by Kaplan-Meier method.Re-sults Immunohistochemical results showed that the strong positive expression rate of SERPINA4 was 97.10%in nor-mal mucosa,71.20%in primary lesions,and 39.40%in metastatic lesions.The difference between the three groups was statistically significant(P<0.001).qRT-PCR results showed that SERPINA4 mRNA was lowly expressed in me-tastatic lesions of metastatic CRC(P<0.05).The expression of SERPINA4 in metastatic lesions was significantly cor-related with histological grade,N stage and M stage of the tumor(P<0.05),but not significantly correlated with age,gender,tumor location and T stage of patients(P>0.05).The survival time of patients with metastatic colorectal cancer with high expression of SERPINA4 was longer than that of patients with low expression.Conclusion The ex-pression of SERPINA4 is of great significance in the survival and prognosis of patients with metastatic CRC.
10.Three-dimensional radiographic characteristics of 18 odontogenic myxoma
Liuyang QU ; Xiaotong LING ; Danni ZHENG ; Jing YANG ; Xuebing YAN ; Denggao LIU ; Yan GAO
Journal of Practical Stomatology 2025;41(5):680-684
Objective:To analyze the there-dimensional radiographic characteristics of odontogenic myxoma(OM).Methods:Clinical records,histopathological reports,SCT or CBCT images of 18 patients with OM were retrospectively collected,and radiograph-ic features including lesion site,shape,size,expansion,intralesional septa and relationship with surrounding tissues were analyzed.The lesions were classified into types by the forms of septa.Results:18 patients(8 males and 10 females,aged 9-39 years old)were included.Among them,12 lesions(66.7%)were round-like in shape,7(38.9%)were located in the posterior maxilla,and 8(44.4%)in the posterior mandible.According to the septa map,the lesions included 4(22.2%)without septum(unilocular type),4 with(22.2%)sporadic septum on the periphery of the tumor(peripheral type),4(22.2%)with the septum from the periphery towards the center of the tumor(centripetal type)and 6(16.7%)with multiple septa interwoven as grid-like trabeclations(criss-crossing tup).Labial/buccal cortex expansion was present in 11 lesions(61.1%),and palatal cortex expansion in 12 lesions(66.7%).Moreover,cortex discontinuity occurred in 14 lesions(77.8%),impacted tooth was present in 9 lesions(16.7%),and root resorption was present in 8 lesions(44.4%).Conclusion:Most OMs occur in the posterior jaw,with multilocular and discontinued cortex.OMs can be classified into unilocular,perpheral,centripetal and cirss-crossing type by the forms of septa in the lesion.

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