1.Transcriptomic responses of Bulinus globosus to extreme temperature and drought stress
Xinyao WANG ; Dandan PENG ; Ying YANG ; Jianfeng ZHANG ; Zhiqiang QIN ; Kun YANG ; Shizhu LI ; Jing XU
Chinese Journal of Schistosomiasis Control 2026;38(1):29-37
Objective To examine the impact of extreme temperature and drought stress on the survival of Bulinus globosus, so as to provide the theoretical evidence for the genomic research of Bulinus in absence of reference genes. Methods B. globosus snail samples were collected from Kiwani Shehia in Pemba Island, Zanzibar, Tanzania, and offspring snails were obtained through laboratory breeding and reproduction. A total of 120 10-week-old B. globosus snails from the same generation were selected and randomly assigned into four groups, including the high-temperature drought (HD) group, normal temperature drought (D) group, low-temperature drought (LD) group, and the control (C) group, of 30 snails in each group. Snails in HD, D, and LD groups were placed in beakers containing dry soil at the bottom and subsequently housed in climate chambers at 35, 26 ℃, and 10 ℃, respectively, while snails in Group C were maintained in 500 mL petri dishes containing dechlorinated tap water at 26 ℃. Following 3 days of breeding, living snails in each group were collected, and soft tissues were dissected and isolated. Total RNA was extracted from snail soft tissues for library construction, followed by high-throughput sequencing on the Illumina HiSeq 4000 sequencing system. De novo transcriptome assembly was performed using the Trinity software, and the longest transcripts were selected as unigenes. Gene functional annotations of unigenes were conducted using the Diamond software against Gene Ontology (GO) knowledgebase, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, NCBI non-redundant (NR) protein sequences database, Protein Family (Pfam) database, and UniProtKB/Swiss-Prot (Swiss-Prot) knowledgebase. GO and KEGG enrichment analyses of differentially expressed genes (DEGs) were performed using the topGO and clusterProfiler software, respectively. In addition, four relevant genes were selected for validation using a real-time quantitative PCR (qRT-PCR) assay to verify the reliability of transcriptome sequencing results. Results Following 3 days of breeding, there were 7, 20, 28, and 30 survival B. globosus snails in HD, LD, D, and C groups, with corresponding survival rates of 23.33% (7/30), 66.67% (20/30), 93.33% (28/30), and 100.00% (30/30), respectively (χ2 = 52.72, P < 0.001). De novo transcriptome assembly generated 176 942 unigenes, with annotation rates of 0.98%, 13.49%, 26.46%, 12.48%, and 14.39% against GO knowledgebase, KEGG pathway database, NR protein sequences database, Pfam database, and Swiss-Prot knowledgebase, respectively. There were 33 up-regulated and 72 down-regulated genes in Group D, 483 up-regulated and 815 down-regulated genes in Group HD, and 245 up-regulated and 172 down-regulated genes in Group LD relative to in Group C. Following removal of overlapping genes across groups and unmatched genes, 11 candidate genes were identified. GO and KEGG analyses revealed 3 heat shock protein (HSP)-related DEGs in these 11 candidate genes, which were annotated as HSP12.2, HSP70, and HSP20 genes and were all significantly up-regulated in each treatment group. Three immune and nervous system-related DEGs were identified, and were all significantly down-regulated in each treatment group, which were involved in the neural cell adhesion molecule L1-like protein pathway, fibrinogen binding protein pathway, and leukocyte elastase inhibitor-like protein pathway. qRT-PCR assay quantified that the expression trends of four genes related to temperature and drought stress across different treatment groups were highly consistent with transcriptome sequencing data. Conclusion The survival rate of B. globosus significantly reduces under combined stresses of extreme temperature and drought, possibly due to an imbalance in its cellular homeostasis regulatory system.
2.Differences in arousal threshold among obstructive sleep apnea patients of different genetic backgrounds and influencing factors
Rui ZHAO ; Ping YAO ; Zhiqiang ZHANG ; Zhiguo GUO ; Minqi XIE ; Hui DANG ; Yanrong JIA ; Jing CHENG ; Dongsheng LYU
Sichuan Mental Health 2026;39(3):240-245
BackgroundObstructive sleep apnea (OSA) represents a prevalent sleep disordered breathing condition characterized by complex pathophysiology. The arousal threshold (ArTH), a core non-anatomical contributor to OSA pathogenesis, is intimately tied to the disease severity and clinical phenotypes. To date, research regarding factors associated with ArTH has yielded inconsistent findings, and ArTH profiles and disparities across populations with different genetic backgrounds are not fully elucidated. ObjectiveTo explore the differences of ArTH in OSA patients with different genetic backgrounds and analyze the key factors affecting ArTH, thereby providing evidence for understanding OSA pathophysiology and formulating targeted treatment regimens. MethodsA total of 285 patients who met the diagnostic criteria for OSA, as defined by the Multidisciplinary Diagnosis and Treatment Guidelines for Adult Obstructive Sleep Apnea, were retrospectively enrolled in this study. All participants underwent overnight polysomnography (PSG) at the Sleep Medicine Center of Inner Mongolia Mental Health Center from December 2022 to May 2024. Based on the study design, the cohort was stratified into two distinct genetic background subgroups (group A and group B). Demographic and clinical characteristics, the Epworth Sleepiness Scale (ESS) score, and overnight PSG data were collected. The apnea hypopnea index (AHI), the lowest pulse oxygen saturation (LSpO2), and fraction of hypopneas (FHypopneas) were utilized as surrogate indicators to estimate ArTH in OSA patients. The influencing factors of low ArTH were tested by binary Logistic regression analysis. ResultsAmong the 285 OSA patients, there were 227 cases (79.65%) in group A and 58 cases (20.35%) in group B. Comparisons between the two genetic background subgroups revealed no statistically significant differences in the proportion of low ArTH, ESS score, PSG parameters, and the three markers for low ArTH (AHI<30 events/h, LSpO2>82.5%, FHypopneas>58.3%) (P>0.05). Binary Logistic regression analysis identified sex (OR=2.421, 95% CI: 1.070–5.478), BMI (OR=0.847, 95% CI: 0.770–0.932), N1 sleep duration (OR=0.974, 95% CI: 0.963–0.985), and hypertension (OR=0.348, 95% CI: 0.143–0.848) as independent factors associated with low ArTH in the total cohort. Stratified analysis by genetic backgrounds revealed that sex(OR=3.799, 95% CI: 1.389–10.392), BMI(OR=0.819, 95% CI:0.723–0.929), and N1 sleep duration(OR=0.973, 95% CI: 0.961–0.986) were independent factors of low ArTH in group A. In contrast, only N1 sleep duration (OR=0.951, 95% CI: 0.911–0.993) remained a significant factor in group B. ConclusionArthur may have conservative characteristics in patients with OSA with different genetic backgrounds, but the pathophysiological mechanism of OSA may have population heterogeneity. [Funded by Inner Mongolia Autonomous Region Natural Science Fundation Project (number, 2024QN08050); Intra-institutional Scientific Research Project of Inner Mongolia Mental Health Center (number, 2022QNWN0010)]
3.Effects of Xiaomudan Granules on PERK/eIF2α/ATF4 pathway and lipid metabolism in mice with nonalcoholic fatty liver
Haitao LIU ; Jingtao LI ; Longmei LI ; Qingrui YANG ; Xinzhu LI ; Yuyu LEI ; Zhiqiang JING
International Journal of Traditional Chinese Medicine 2025;47(7):932-937
Objective:To investigate the effects of Xiaomudan Granules on PERK/eIF2α/ATF4 pathway and lipid metabolism in NAFLD mice.Methods:Totally 60 mice were divided into a normal group of 10 mice and a high-fat diet group of 50 mice. The high-fat diet group was given a high-fat diet to establish a NAFLD mouse model. The high-fat feed group was divided into model group, metformin group, and Xiaomudan Granules low-, medium-, and high-dosage groups according to the random number table method. Xiaomudan Granules low-, medium-, and high-dosage groups were orally administered with concentrated Xiaomudan Granules at dosages of 11.7, 23.4, and 46.8 g/kg, respectively. The metformin group was orally administered with metformin solution at a dose of 0.2 g/kg, while the normal group and model group were orally administered with distilled water of equal volume once a day for 8 consecutive weeks. The liver oil red O staining of mice was observed in each group. The protein expressions of p-PERK, p-eIF2α, ATF4, C/EBP homologous protein (CHOP), CCAAT enhancer binding protein alpha (C/EBPα), C/EBPβ, and steroid regulatory element binding protein 2 (SREBP2) were detected by Western blot. Fluorescence quantitative PCR was used to detect the mRNA expressions of ATF4, CHOP, C/EBPα, C/EBPβ, and SREBP2.Results:The liver tissue structure and steatosis of mice were improved in Xiaomudan Granules groups. Compared with the model group, the expressions of p-PERK/PERK, ATF4, C/EBPα, C/EBPβ, and SREBP2 proteins decreased in Xiaomudan Granules groups and the metformin group ( P<0.01 or P<0.05). The expressions of p-eIF2α/eIF2α and CHOP proteins decreased in the Xiaomudan Granules medium- and high-dosage groups and the metformin group ( P<0.01); compared with the model group, the levels of ATF4, CHOP, C/EBPα, and C/EBPβ mRNA in Xiaomudan Granules groups and the metformin group decreased ( P<0.01), while the levels of SREBP2 mRNA in the Xiaomudan Granules medium- and high-dosage groups and the metformin group decreased ( P<0.01). Conclusion:Xiaomudan Granules may improve liver lipid metabolism and reduce liver fat deposition in NAFLD mice by regulating the PERK/eIF2α/ATF4 pathway.
4.Preliminary clinical study of miniprobe echoendoscope combined with balloon dilation in the treatment of refractory benign esophageal stenosis
Jing TANG ; Zhiqiang YI ; Qiaomu LUO ; Tao WU ; Dan YANG ; Jing KUANG ; Aimin LIU
China Journal of Endoscopy 2025;31(3):1-6
Objective To explore the clinical value of mini-incision combined with balloon dilatation under the guidance of miniprobe echoendoscope in the treatment of refractory benign esophageal stenosis.Methods From October 2019 to October 2023,30 cases of esophageal early cancer with esophageal scar stenosis after endoscopic submucosal dissection(ESD)were selected and randomly divided into study group(miniprobe echoendoscope guided small incision+balloon dilatation,15 cases)and control group[esophageal radial incision(ERI),15 cases],EUS in the study group judged the hierarchical structure of esophageal wall before operation,and excluded those with esophageal cancer recurrence and submucosal invasive growth.According to miniprobe echoendoscope,the length of scar segment and the scope of scar ring lumen were defined,and the location and length of pre-incision were defined,and then balloon was used to expand step by step.The short-term postoperative complications,Stooler dysphagia scale score and restenosis rate of the two groups were observed,and the feasibility,safety and efficacy were evaluated.Results The incidence of short-term postoperative complications 6.7%in the study group was significantly lower than that 46.7%in the control group,the difference was statistically significant(P<0.05),and the Stooler dysphagia scale score were(1.133±0.990)and(1.600±0.737)in the study group at 1 and 2 months after operation with significantly lower than those(2.067±1.033)and(2.467±0.915)in the control group,the differences were statistically significant(P<0.05).Two months after operation,the restenosis rate was 13.3%,which was significantly lower than that 46.7%of the control group,the difference was statistically significant(P<0.05),and the scar thickness under miniprobe echoendoscope(2.113±0.887)mm was significantly thinner than that(3.353±1.468)mm of the control group,the difference was statistically significant(P<0.05).Conclusion Preoperative miniprobe echoendoscope evaluation,can accurately locate lesions,endoscopic mini-incision combined with balloon mechanical expansion are safe and effective,with less pain and fewer postoperative complication.
5.Reliability and validity test of the Chinese version of the Urinary Incontinence Awareness and Attitude Scale
Zhiqiang CHENG ; Baozhen ZHANG ; Liping TANG ; Jing LI ; Jiaoyun XIA ; Xueyan WEI ; Zhixian GONG ; Meizhen ZHANG ; Lusi LI
Chinese Journal of Nursing 2025;60(9):1107-1112
Objective To translate the Urinary Incontinence Awareness and Attitude Scale(URINAS)and test its reliability and validity.Methods The Chinese version of the URINAS was developed by Brislin's translation model for translation,back translation,cultural adaptation,and pilot investigation.A convenience sampling method was used to select 384 urinary incontinence patients who visited a tertiary hospital in Nanchang,Jiangxi Province from June 2024 to October 2024 for investigation,in order to evaluate the reliability and validity of the scale.Results The Chinese version of the URINAS consisted of 5 dimensions with 26 entries.The Cronbach's alpha coefficient of this scale was 0.843;the folded half reliability was 0.917;the retest reliability was 0.852.The content validity of the scale at the level of the entries ranged from 0.846 to 1.000,and that at the level of the scale was 0.979.A total of 5 metrics were extracted by exploratory factor analysis,and the cumulative variance contribution rate of 74.286%.The results of the validation factor analysis showed a chi-square/degree of freedom of 2.268,a root mean square error of approximation of 0.064,a standardized fit index of 0.916,a Tucker-Lewis index of 0.923,a comparative fit index of 0.906,and a goodness-of-fit index of 0.922.Conclusion The URINAS has good reliability and validity,and can better reflect the level of patients' cognition and attitude towards urinary incontinence,thus providing theoretical basis for the development of corresponding intervention programs for such patients.
6.Predictive value of caliceal pelvic height-to-infundibular length ratio for stone-free rate in lower calyx stone treatment with flexible ureteroscopic lithotripsy
Shiwei HUANG ; Sheng ZHONG ; Guangming YIN ; Long WANG ; Zhiqiang JIANG ; Kai HUANG ; Jing TAN
Chinese Journal of Urology 2025;46(8):600-606
Objective:This study aims to explore the predictive value of the ratio of caliceal pelvic height to infundibular length(CPH/IL)for the stone-free rate(SFR)in the treatment of lower calyx stones using flexible ureteroscopic lithotripsy(FURL)combined with a distally bendable negative pressure suction sheath.Methods:A retrospective analysis was conducted on the clinical data and anatomical parameters of 312 patients with lower calyx stones or combined lower calyx stones admitted to the Third Xiangya Hospital of Central South University from September 2022 to December 2023,all of whom were treated with FURL combined with a distally bendable negative pressure suction sheath. Stone clearance was defined as no residual stones or residual stones with a diameter of ≤3 mm without any symptoms. Patients were divided into the clearance group(265 cases,84.90%)and the non-clearance group(47 cases,15.10%). There were no significant differences in gender(male/female:173/92 cases vs. 29/18 cases),age[(44.69 ± 13.14)years vs.(42.60 ± 10.93)years],degree of hydronephrosis(no hydronephrosis/mild hydronephrosis/moderate to severe hydronephrosis:122/85/58 cases vs. 21/12/14 cases),side of lower calyx stones(left/right:157/108 cases vs. 31/16 cases),maximum diameter of stones(MDS)[(19.23 ± 6.41)mm vs.(17.77 ± 6.18)mm],and CT value of stones[(993.46 ± 249.12)Hu vs.(1013.43 ± 300.90)Hu]between the two groups( P > 0.05),indicating comparability between groups. There was no significant difference in the distance from the midpoint of the lower lip of the renal pelvis to the ureter at the lowest plane of the lower calyx(K-A line)between the clearance and non-clearance groups[(26.16 ± 5.18)mm vs.(25.70 ± 8.66)mm, P > 0.05]. However,significant differences were observed in the infundibulopelvic angle(IPA)[(53.97 ± 15.72)° vs.(37.43 ± 15.39)°],infundibular length(IL)[(27.26 ± 5.11)mm vs.(33.04 ± 7.38)mm],infundibular width(IW)[(8.27 ± 2.82)mm vs.(7.09 ± 3.20)mm],caliceal pelvic height(CPH)[(19.96 ± 4.63)mm vs.(30.32 ± 7.56)mm],ureter-lower calyx distance(ULD)[(23.00 ± 5.59)mm vs.(18.78 ± 6.31)mm],CPH/IL ratio[(0.73 ± 0.11)vs.(0.92 ± 0.09)],and curvature of the lower calyx of the renal pelvis[(0.06 ± 0.01)mm -1 vs.(0.08 ± 0.03)mm -1]between the two groups( P <0.05). Univariate and logistic multivariate regression analyses were used to identify the independent risk factors affecting the postoperative SFR of FURL-treated lower calyx stones and to assess the value of CPH/IL for SFR in the treatment of lower calyx stones using FURL combined with a distally bendable negative pressure suction sheath. A logistic multivariate regression model and a corresponding nomogram were constructed,and the predictive ability of the model for SFR was evaluated using the receiver operating characteristic(ROC)curve. The calibration curve and the Hosmer -Lemeshow test were used to assess the consistency and accuracy of the model. The clinical utility of the model was evaluated using decision curve analysis(DCA). Results:Univariate analysis revealed that seven anatomical parameters differed significantly between the clearance and non-clearance groups(all P < 0.05):infundibulopelvic angle(IPA, OR = 0.27,95% CI 0.17-0.42, P < 0.01),infundibular length(IL, OR = 2.52,95% CI 1.83-3.47, P < 0.01),infundibular width(IW, OR = 0.64,95% CI 0.45-0.90, P = 0.011),caliceal pelvic height(CPH, OR = 5.78,95% CI 3.67-9.10, P < 0.01),ureter-lower calyx distance(ULD, OR = 0.43,95% CI 0.30-0.63, P < 0.01),CPH/IL ratio( OR = 13.62,95% CI 6.86-27.03, P < 0.01),and curvature of the lower calyx of the renal pelvis( O = 3.15,95% CI 2.08-4.78, P < 0.01). Multivariate logistic regression further identified CPH/IL( OR = 9.87,95% CI 4.92-19.79, P < 0.01),IPA( OR = 0.41,95% CI 0.23-0.71, P = 0.001),and ULD( OR = 0.50,95% CI 0.29-0.87, P = 0.014)as independent risk factors influencing the stone-free rate after flexible ureteroscopic lithotripsy combined with a distally bendable negative-pressure suction sheath. The area under the ROC curve(AUC)of the univariate regression model showed that for IPA,the AUC was 0.788(95% CI 0.711-0.864);for IL,it was 0.731(95% CI 0.643-0.819);for ULD,it was 0.677(95% CI 0.586-0.767);for CPH,it was 0.867(95% CI 0.804-0.929);for IW,it was 0.628(95% CI 0.535-0.721);for CPH/IL,it was 0.906(95% CI 0.850-0.961)with an optimal cutoff value of 0.882,corresponding to a sensitivity of 83.02% and a specificity of 92.08%;and for curvature of the lower calyx of the renal pelvis,it was 0.744(95% CI 0.662-0.827). The AUC of the multivariate regression model was 0.929(95% CI 0.884-0.974)with an optimal cutoff value of 0.364,corresponding to a sensitivity of 82.98% and a specificity of 95.09%. Calibration curves demonstrated close agreement between the predicted and actual stone-free rates after FURL(C-index = 0.921). The Hosmer-Lemeshow goodness-of-fit test indicated no significant discrepancy between predicted and observed probabilities( P = 0.135,>0.05). Decision-curve analysis further revealed that basing clinical decisions on the model ,s predicted probability yields a higher net clinical benefit than either withholding FURL for all patients or treating all patients regardless of risk,and it also outperforms strategies guided by any single independent predictor alone. Conclusions:IPA,IL,IW,CPH,ULD,CPH/IL ratio,and curvature of the lower calyx of the renal pelvis can all predict the postoperative SFR of FURL combined with a distally bendable negative pressure suction sheath for the treatment of lower calyx stones to varying degrees. Among them,CPH/IL has the highest predictive value. When CPH/IL >0.88,it may be difficult to use FURL to treat lower calyx stones.
7.Role effectiveness and skill application of simulated participants in medical scenario simulation- based teaching
Wenlong YAO ; Li LI ; Jing XIAO ; Zhiqiang ZHOU ; Chang ZHU ; Li WAN
Chinese Journal of Medical Education Research 2025;24(6):771-776
Objective:To investigate the roles of simulated participants in the facilitation of medical scenarios simulation-based course and to provide a reference for the standardized construction of medical scenarios simulation-based course.Methods:Thirty scenario simulation-based teaching sessions on perioperative crisis events conducted during the 2022-2023 academic year were observed on-site. The performance of simulated participants in facilitating case execution and the skills used in facilitation were observed and recorded. At the end of the course, surveys regarding facilitation were administered to students, simulated participants, and instructors for analysis.Results:In the facilitation of 30 scenario simulation-based teaching sessions on perioperative crisis events, the roles of simulated participants included surgeon, anesthesia nurse, circuit nurse, and standardized patient. Two simulated participants accounted for 80.00% and three participants for 20.00% of the total sessions. The simulated participants performed excellently in helping the learners quickly integrate into the scenario and controlling the direction of the case, with an excellence rate of 90.00%-100.00%. However, they had some variations in driving the progress of the case and responding to unexpected situations, with an excellence rate of 50.00%-60.00%. The facilitation skills used by simulated participants included hinting, prompting, and instructing. The performance of facilitation was closely related to the use of skills by the simulated participants. Simulated participants with proficient skill usage significantly enhanced teaching effectiveness. Post-session surveys revealed that learners generally perceived simulated participants as having a positive impact on facilitation, and both simulated participants and instructors emphasized the need for strengthened communication prior to simulation-based teaching.Conclusions:The simulated participants played important roles in guiding learners, advancing case progression, and achieving teaching objectives. Instructors should not only consider the scripting of simulated participants during lesson planning but also prioritize their training and communication during course implementation.
8.Research on the association of breast cancer patients'prognosis with internal mammary sentinel lymph node biopsy
Ruijie JIA ; Zhiqiang SHI ; Qi ZHANG ; Yongjin LU ; Junsheng ZHENG ; Jing SUN ; Zhao BI ; Xiao SUN ; Yongsheng WANG ; Pengfei QIU
China Oncology 2025;35(4):394-403
Background and purpose:Internal mammary sentinel lymph node biopsy(IMSLNB)is a minimally invasive diagnostic technique for regional lymph nodes in breast cancer,which can provide accurate lymph staging and guide adjuvant treatment decision,but its clinical application has been controversial.The purpose of this study was to investigate the prognosis of IMSLNB in early breast cancer.Methods:In this study,a retrospective cohort of 7 949 patients with breast cancer from January 1,2016 to December 31,2021 was analyzed.After applying propensity score matching,the patients were divided into IMSLNB group and no-IMSLNB group,and the regional recurrence-free survival(RRFS),local recurrence-free survival(LRFS),distant metastasis-free survival(DMFS),disease-free survival(DFS)and overall survival(OS)of the two groups were compared.This study was approved by the Clinical Research Ethics Committee of the Affiliated Tumor Hospital of Shandong First Medical University(approval number:SDTHEC20130324).Results:A total of 990 patients were included in the final analysis(330 in the IMSLNB group and 660 in the no-IMSLNB group).IMSLN metastasis was found in 54 patients in the IMSLNB group,including 47 patients with axillary lymph node(ALN)metastasis and 7 patients with IMSLN metastasis only.The IMSLN transfer rate was 16.4%.The median follow-up of 41 months showed that the IMSLNB group demonstrated better 3-year DFS[98.4%vs 94.2%,hazard ratio(HR)=0.509;95%CI:0.312-0.828,P=0.007]and 5-year DFS(92.5%vs 87.3%,HR=0.214,95%CI:0.206-0.222,P=0.011)compared with no-IMSLNB group.However,no significant differences were observed in 3-year OS(99.1%vs 99.4%,HR=0.618,95%CI:0.231-1.655,P=0.338)or 5-year OS(98.5%vs 99.1%,HR=0.52,95%CI:0.51-0.53,P=0.392)between the two groups.The 3-year RRFS in the IMSLNB group was better compared with the no-IMSLNB group(99.09%vs 97.73%,HR=0.066;95%CI:0.061-0.071,P=0.048),while no significant differences were observed in 3-year LRFS(99.70%vs 98.19%,HR=0.209;95%CI:0.201-0.217,P=0.130)or DMFS(95.76%vs 96.06%,HR=0.865,95%CI:0.858-0.872,P=0.820)between the two groups.The exploratory subgroup analysis of DFS revealed that patients in the following subgroups could significantly benefit from IM-SLNB(P<0.05):diagnosis age(≤50 years),premenopausal status,BMI(≤24),lymphovascular invasion(LVI,present),tumor location(lateral),molecular subtype[hormone receptor positive(HR+)/human epidermal growth factor receptor 2 negative(HER2-)],histological type(invasive ductal carcinoma),and axillary lymph node status(positive).Conclusion:IMSLNB can provide more accurate regional lymph node staging for early breast cancer,help optimize adjuvant radiotherapy strategies,and improve patients'DFS and RRFS.It can be promoted as a minimally invasive staging technique for regional lymph nodes.
9.Effect of over-expression of NR2F2 on biological behaviors of human ovarian cancer SKOV3 cells
Shuo ZHANG ; Yunxiu XIA ; Weiwei CHEN ; Hongliang DONG ; Bingjie CUI ; Cuilan LIU ; Zhiqiang LIU ; Fei WANG ; Jing DU
Journal of Jilin University(Medicine Edition) 2025;51(1):58-67
Objective:To investigate the effect of nuclear receptor subfamily 2 group F member 2(NR2F2)on the biological behaviors of human ovarian cancer SKOV3 cells,and to clarify its molecular mechauism and provide the new idea for treatment of ovarian cancer.Methods:Gene Expression Profiling Interactive Analysis(GEPIA)Database analyse the expression level of NR2F2 gene in ovarian tissue,and analyse its correlation with clinical prognosis of ovarian cancer patients.The human ovarian cancer SKOV3 cells were divided into control group and NR2F2 over-expression(NR2F2 OE)group,which were transfected with mCherry control virus and NR2F2 OE over-expression virus,respectively,when the cell deusity reached 70%,and the stable transfection SKOV3 cell lines were screened with puromycin(puro)48h lafter.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the transfection efficiencies of the cells;RT-qPCR method was used to detect the expression levels of NR2F2 and sex-determining region Y-box 2(SOX2)mRNA in the cells in two groups;Western blotting method was used to detect the expression levels of NR2F2,ATP-binding cassette superfamily G member 2(ABCG2),and programmed cell death 1-ligand 1(PD-L1)protcins in the cells in two groups.CCK-8 assay was used to detect the proliferation activities of the cells in two groups;Wound assay was used to detect the migration rates of the cells in two groups;Transwell chamber assay was used to detect the number of transmembrane cells;Spheroidization assay was used to detect the numbers of spheroids in the cells;peripheral blood mononuclear cells(PBMCs)-mediated tumor cell killing assay was used to detect the relative densities of surviving tumor cells;CCK-8 assay was used to detect the half maximal inhibitory concentration(IC50)of paclitaxel(PTX)and carboplatin(CBP).Results:Compared with normal ovarian tissue,the expression level of NR2F2 gene in ovarian tumor tissue was decreased(P<0.05),and decreased with the improvement of clinical pathological grading of ovarian tumor.The patients with higher expression level of NR2F2 gene had better clincal prognosis.The SKOV3 cells with NR2F2 over-expresson were successfully constructed,and the expression levels of NR2F2 mRNA and protein in the cells in NR2F2 OE group were increased compared with control group(P<0.001).The CCK-8 assay results showed that compared with control group,the proliferation activities of the cells in NR2F2 OE group were decreased at different time points(1,2,3,and 4 d)(P<0.05 or P<0.01).The cell wound assay results showed that compared with control group,the migration rate of the cells in NR2F2 OE group was decreased(P<0.001).The Transwell assay results showed that compared with control group,the number of transmembrane cells in NR2F2 OE group was decreased(P<0.01).Compared with control group,the number of the spheroids in NR2F2 OE group was decreased(P<0.05),and the expression levels of SOX2 mRNA(P<0.01)and protein(P<0.001)were increased.Compared with control group,the relative density of surviving tumor cells in NR2F2 OE group was decreased,but the difference was not significant(P<0.05),and the expression level of PD-L1 protein was decreased(P<0.05).Compared with control group,the proliferation activities of cells in NR2F2 OE group were decreased(P<0.05),and the drug sensitivities of the cells to PTX and CBP were enhanced(P<0.05);the IC50 of PTX was significantly reduced,while the IC50of CBP could not be calculated due to excessively high drug concentration;the expression level of ABCG2 protein was decreased(P<0.05).Conclusion:The over-expression of NR2F2 may inhibit the proliferation,migration,and invasion of the human ovarian cancer SKOV3 cells,decrease the expression levels of SOX2,PD-L1 and ABCG2 proteins,suppress the stemness and immune evasion ability of the SKOV3 cells,and enhance the sensitivities of the SKOV3 cells to PTX and CBP.
10.Preliminary clinical exploration of endoscopic ultrasound combined with modified endoscopic mucosal resection in the treatment of rectal neuroendocrine tumors
Ping LUO ; Aimin LIU ; Zhiqiang YI ; Qiaomu LUO ; Sha WEI ; Jing KUANG ; Jing TANG
Chongqing Medicine 2025;54(4):893-897
Objective To investigate the clinical feasibility and safety of endoscopic ultrasonography(EUS)combined with modified endoscopic mucosal resection(EMR)in the treatment of rectal neuroendo-crine tumors(RNETs).Methods A total of 48 patients diagnosed with RNETs by colonoscopy in the depart-ment of gastroenterology in this hospital from December 2021 to June 2023 were selected as the study objects.Patients were randomly divided into the study group(EUS combined with modified EMR,n=16),the control 1 group(traditional EMR,n=16)and the control 2 group[endoscopic submucosal dissection(ESD),n=16].The operation time,R0 resection rate and postoperative complications of each group were observed.Endoscop-ic ultrasonography was followed up 3 and 6 months after surgery to determine whether there was any recur-rence.Results The operative time of the study group[(17.813±0.379)min]was significantly shorter than that of the control 2 group[(36.250±3.296)min],the difference was statistically significant(P<0.05),but compared with the control 1 group[(16.375±1.996)min],there was no significant difference(P>0.05).The incidence of complications in the study group(6.2%)was significantly lower than that in the control 2 group(37.5%),the difference was statistically significant(P<0.05).while the incidence of complications in the study group was not significantly higher than that in the control 1 group(12.5%,P>0.05).R0 removal rate in the study group(93.8%)was significantly higher than that in the control 1 group(62.5%)and the control 2 group(75.0%),the difference was statistically significant(P<0.05).Conclusion EUS combined with modified EMR has more advantages than EMR and ESD in the treatment of RNETs,and has certain fea-sibility and safety,which is convenient for clinical application.

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