1.Mechanism of piRNA-2732 Promoting Proliferation,Migration and Invasion of Cervical Cancer Cells through METTL3 Mediated m6A RNA Methylation
Miaomiao LIU ; Shuangshuang XIE ; Wei LI ; Jinyi WANG ; Yueyue GAO ; Yanhua KANG
Journal of Modern Laboratory Medicine 2025;40(5):22-27
Objective To explore the biological functions and mechanisms of PIWI-interacting RNA(piRNA)in cervical cancer(CC).Methods RT-qPCR was used to detect the expression level of piRNA-2732 in CC tissue,CaSki cells and End1/E6E7 cells.EpiQuik m6A RNA methylation quantification kit was used to detect the methylation level of m6A RNA in CaSki cells.The expression levels of methyltransferases(METTL3,METTL14 and WTAP)and demethylases(FTO,ALKBH5)mRNA in CaSki cells were detected by RT-qPCR.After culturing CaSki cells to logarithmic growth stage,they were divided into six groups:piRNA-2732 mimic negative control group(mi-NC group),piRNA-2732 mimic group(mi-2732 group),piRNA-2732 inhibitor negative control group(in-NC group),piRNA-2732 inhibitor group(in-2732 group),piRNA-2732 mimic+METTL3 knockdown control group(mi-2732+si-NC group),and piRNA-2732 mimic+METTL3 knockdown group(mi-2732+si-METTL3 group).The viability of CaSki cells was detected by CCK8 assay.Colony formation assay was used to detect the proliferation ability of CaSki cells.Transwell experiment was used to detect the migration and invasion ability of CaSki cells.RT-qPCR and Western blot were used to detect the expression of methyltransferase like protein 3(METTL3).Transfected METTL3 wild-type(METTL3-WT)and METTL3 mutant(METTL3-MUT)in the mi-NC group,mi-2732 group,in-NC group,and in-2732 group respectively,and detected the effect of piRNA-2732 on METTL3 through dual luciferase reporter gene assay.Results Compared with the adjacent tissues,the expression of piRNA-2732(3.84±1.08 vs 1.32±0.53)was significantly higher in the cancer tissues of CC patients,and the difference was statistically significant(t=5.115,P<0.001).Compared with end1/E6E7 cells,the expression of piRNA-2732(1.00±0.13 vs 1.67±0.16)in CaSki cells was significantly higher,and the difference was statistically significant(t=5.632,P<0.01).Compared with mi-NC group,mi-2732 group promoted the viability,proliferation,migration and invasion of CaSki cells,and the differences were statistically significant(t=4.410~11.040,all P<0.01).Compared with mi-NC group,mi-2732 group increased m6A RNA methylation level and METTL3 mRNA and protein,the differences were statistically significant(t=6.176,9.211,12.550,all P<0.05).The results of dual luciferase reporter gene testing showed that compared with the mi-NC+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-WT group was significantly increased(t=11.850).Compared with mi-2732+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-MUT group was significantly lower(t=12.740),and the difference was statistically significant(all P<0.000 1).Compared with in NC+METTL3-WT group,the relative luciferase activity of in-2732+METTL3-WT group was significantly lower(t=7.828),compared with in-2732+METTL3-WT group,the relative luciferase activity of CaSki cells in in-2732+METTL3-MUT group was significantly increased(t=8.146),and the difference was statistically significant(all P<0.001).Compared with mi-2732+si-NC group,the expression level of m6A in mi-2732+si-METTL3 group was significantly lower,and the difference was statistically significant(t=7.630,P<0.01).Compared with mi-2732+si-NC group,the proliferation ability,colony number,cell migration and invasion ability of CaSki cells in mi-2732+si-METTL3 group were significantly decreased,and the differences were statistically significant(t=3.695~4.891,all P<0.001).Conclusion piRNA-2732 is overexpressed in CC tissues and cells,and piRNA-2732 promotes tumor development in CC through METTL3 mediated m6A methylation.
2.Qualitative study on pain experiences in patients with post-stroke hemiplegic shoulder pain based on the IKAP theory
Yifan KANG ; Li WANG ; Xinyi ZHANG ; Menglin YUAN ; Miaomiao ZHAO ; Shuangshuang DU
Chinese Journal of Practical Nursing 2025;41(17):1309-1315
Objective:To explore the pain sensations and experiences of patients with post-stroke hemiplegic shoulder pain, and to provide reference for the formulation of intervention plans.Methods:Objective sampling method was used to conduct semi-structured interviews on 15 patients with post-stroke hemiplegic shoulder pain in the department of neurology and rehabilitation department of Wuhan No.1 Hospital from January to March 2024. The interview outline was established based on the information-knowledge-attitude-practice (IKAP) theory, and the data were analyzed, summarized and extracted by Colaizzi 7-step analysis method.Results:According to the four core components of the IKAP theory, ten themes were extracted. The information and knowledge included the neglect of the importance of early management for hemiplegic shoulder pain, the lack of understanding of the relevant knowledge about hemiplegic shoulder pain, and the use of restricted or incorrect attribution methods in relation to hemiplegic shoulder pain. The attitude included a perception of pain as uncontrollable, the suppression of pain expression, the concern regarding the persistent presence of pain, and the experience of negative emotions. The practice included the adverse effects stemming from hemiplegic shoulder pain, the strategies employed to cope with hemiplegic shoulder pain, and the expectation of multifaceted social support.Conclusions:Patients with post-stroke hemiplegic shoulder pain bear many burdens and have needs in terms of pain experiences. Healthcare professionals need to strengthen pain health education and guidance, help patients develop correct pain cognition and beliefs, refine the pain management system, assist patients in establishing a diverse social support system, enhance their pain coping abilities, and comprehensively promote the rehabilitation of the disease.
3.Mechanism of piRNA-2732 Promoting Proliferation,Migration and Invasion of Cervical Cancer Cells through METTL3 Mediated m6A RNA Methylation
Miaomiao LIU ; Shuangshuang XIE ; Wei LI ; Jinyi WANG ; Yueyue GAO ; Yanhua KANG
Journal of Modern Laboratory Medicine 2025;40(5):22-27
Objective To explore the biological functions and mechanisms of PIWI-interacting RNA(piRNA)in cervical cancer(CC).Methods RT-qPCR was used to detect the expression level of piRNA-2732 in CC tissue,CaSki cells and End1/E6E7 cells.EpiQuik m6A RNA methylation quantification kit was used to detect the methylation level of m6A RNA in CaSki cells.The expression levels of methyltransferases(METTL3,METTL14 and WTAP)and demethylases(FTO,ALKBH5)mRNA in CaSki cells were detected by RT-qPCR.After culturing CaSki cells to logarithmic growth stage,they were divided into six groups:piRNA-2732 mimic negative control group(mi-NC group),piRNA-2732 mimic group(mi-2732 group),piRNA-2732 inhibitor negative control group(in-NC group),piRNA-2732 inhibitor group(in-2732 group),piRNA-2732 mimic+METTL3 knockdown control group(mi-2732+si-NC group),and piRNA-2732 mimic+METTL3 knockdown group(mi-2732+si-METTL3 group).The viability of CaSki cells was detected by CCK8 assay.Colony formation assay was used to detect the proliferation ability of CaSki cells.Transwell experiment was used to detect the migration and invasion ability of CaSki cells.RT-qPCR and Western blot were used to detect the expression of methyltransferase like protein 3(METTL3).Transfected METTL3 wild-type(METTL3-WT)and METTL3 mutant(METTL3-MUT)in the mi-NC group,mi-2732 group,in-NC group,and in-2732 group respectively,and detected the effect of piRNA-2732 on METTL3 through dual luciferase reporter gene assay.Results Compared with the adjacent tissues,the expression of piRNA-2732(3.84±1.08 vs 1.32±0.53)was significantly higher in the cancer tissues of CC patients,and the difference was statistically significant(t=5.115,P<0.001).Compared with end1/E6E7 cells,the expression of piRNA-2732(1.00±0.13 vs 1.67±0.16)in CaSki cells was significantly higher,and the difference was statistically significant(t=5.632,P<0.01).Compared with mi-NC group,mi-2732 group promoted the viability,proliferation,migration and invasion of CaSki cells,and the differences were statistically significant(t=4.410~11.040,all P<0.01).Compared with mi-NC group,mi-2732 group increased m6A RNA methylation level and METTL3 mRNA and protein,the differences were statistically significant(t=6.176,9.211,12.550,all P<0.05).The results of dual luciferase reporter gene testing showed that compared with the mi-NC+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-WT group was significantly increased(t=11.850).Compared with mi-2732+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-MUT group was significantly lower(t=12.740),and the difference was statistically significant(all P<0.000 1).Compared with in NC+METTL3-WT group,the relative luciferase activity of in-2732+METTL3-WT group was significantly lower(t=7.828),compared with in-2732+METTL3-WT group,the relative luciferase activity of CaSki cells in in-2732+METTL3-MUT group was significantly increased(t=8.146),and the difference was statistically significant(all P<0.001).Compared with mi-2732+si-NC group,the expression level of m6A in mi-2732+si-METTL3 group was significantly lower,and the difference was statistically significant(t=7.630,P<0.01).Compared with mi-2732+si-NC group,the proliferation ability,colony number,cell migration and invasion ability of CaSki cells in mi-2732+si-METTL3 group were significantly decreased,and the differences were statistically significant(t=3.695~4.891,all P<0.001).Conclusion piRNA-2732 is overexpressed in CC tissues and cells,and piRNA-2732 promotes tumor development in CC through METTL3 mediated m6A methylation.
4.Qualitative study on pain experiences in patients with post-stroke hemiplegic shoulder pain based on the IKAP theory
Yifan KANG ; Li WANG ; Xinyi ZHANG ; Menglin YUAN ; Miaomiao ZHAO ; Shuangshuang DU
Chinese Journal of Practical Nursing 2025;41(17):1309-1315
Objective:To explore the pain sensations and experiences of patients with post-stroke hemiplegic shoulder pain, and to provide reference for the formulation of intervention plans.Methods:Objective sampling method was used to conduct semi-structured interviews on 15 patients with post-stroke hemiplegic shoulder pain in the department of neurology and rehabilitation department of Wuhan No.1 Hospital from January to March 2024. The interview outline was established based on the information-knowledge-attitude-practice (IKAP) theory, and the data were analyzed, summarized and extracted by Colaizzi 7-step analysis method.Results:According to the four core components of the IKAP theory, ten themes were extracted. The information and knowledge included the neglect of the importance of early management for hemiplegic shoulder pain, the lack of understanding of the relevant knowledge about hemiplegic shoulder pain, and the use of restricted or incorrect attribution methods in relation to hemiplegic shoulder pain. The attitude included a perception of pain as uncontrollable, the suppression of pain expression, the concern regarding the persistent presence of pain, and the experience of negative emotions. The practice included the adverse effects stemming from hemiplegic shoulder pain, the strategies employed to cope with hemiplegic shoulder pain, and the expectation of multifaceted social support.Conclusions:Patients with post-stroke hemiplegic shoulder pain bear many burdens and have needs in terms of pain experiences. Healthcare professionals need to strengthen pain health education and guidance, help patients develop correct pain cognition and beliefs, refine the pain management system, assist patients in establishing a diverse social support system, enhance their pain coping abilities, and comprehensively promote the rehabilitation of the disease.
5.Comparative study on image quality and radiation dose between the spiral scanning and the axial scanning for skull phantom based on specific conditions
Zhiyuan GAO ; Yan SUI ; Kang LIU ; Zhaorui CHEN ; Shuangshuang ZHENG ; Dewu YANG
China Medical Equipment 2024;21(9):18-22
Objective:To explore and compare the image quality and radiation dose between the spiral scanning and the axial scanning for skull phantom based on specific conditions.Methods:The position of the orbitomeatal base line(OBL)of the skull phantom was marked,and the different angles of elevation of skull were adjusted.The angles between OBL and bed surface were respectively set as 90°,100°,110° and 120°.The axial scanning and spiral scanning were respective adopted to conduct 24 times of image acquisition when the computed tomography dose index(CTDIvol)of fixed volume were respectively 40,50 and 60 mGy.The cross section of axial scanning was vertical to OBL,and the images of spiral scanning used the reconstructed technique of image quality enhancement(IQE)of removing spiral artifacts to conduct reconstruction along the OBL direction.The CT value(HU)and standard deviation(SD)of the bilateral cerebellum,temporal lobe,frontal lobe and parietal lobe of the phantom were measured,and the signal-to-noise ratio(SNR)of each lobe of the phantom was calculated.The CTDIvol and dose-length product(DLP)were recorded,and the effective dose(ED)of lens was calculated.Results:The differences of the SNR values of cerebellum and occipital lobe(R),cerebellum and occipital lobe(L),temporal lobe(R),temporal lobe(L),frontal lobe(R),frontal lobe(L),parietal lobe(R)and parietal lobe(L)between two kinds of scanning models were significant(F=6.48,5.83,7.00,6.20,7.30,8.26,5.72,5.83,P<0.05),respectively.There was significant difference in lens DLP between the two kinds of scanning models(F=10.96,P<0.05).The spiral scanning and IQE reconstructed technique were used to conduct image acquisition and processing.The SNR value of spiral scanning imaging was better than that of axial scanning,and the reconstructions of coronal and sagittal positions could be conducted,and the radiation doses of lens were similar,but the DLP value of spiral scanning was slightly higher than that of axial scanning at the same CTDIvol.Conclusion:Using the IQE reconstruction technique of spiral scanning can obtain satisfactory CT sectional images of skull under the situations that machine cannot change scanning angle or the scanning of axial position is unable to conduct cooperation.
6.Analysis of renal glucose threshold and related factors in patients with type 2 diabetes mellitus
Jinfu SHEN ; Zhuoqun WANG ; Shuangshuang FENG ; Mao LI ; Juan LI ; Tingting GAO ; Jingjing KANG ; Xingpo MA ; Min NIU
Chinese Journal of Endocrinology and Metabolism 2021;37(1):34-38
Objective:To investigate the value of renal glucose threshold and related factors in patients with type 2 diabetes mellitus.Methods:According to the cut-off point of normal renal glucose threshold(RT G 8.9-10 mmol/L), 107 patients with type 2 diabetes mellitus hospitalized in the Endocrinology Department of our hospital were divided into three groups: high RT G group(RT G>10 mmol/L), medium RT G group(8.9 mmol/L≤RT G≤10 mmol/L), and low RT G group(RT G<8.9 mmol/L). The clinical data and biochemical characteristics of each group were collected and analyzed. Results:The proportions of patients with high, medium, and low RT G of type 2 diabetes mellitus were 56%, 29%, and 15%, respectively. There were significant differences in RT G value, age, course of disease, body mass index(BMI), fasting plasma glucose(FPG), HbA 1C, total cholesterol(TC), serum creatinine, mean blood glucose(MBG), and 24-hour urine glucose between high and medium RT G groups. RT G, gender, BMI, FPG, HbA 1C, TC, and MBG in patients with high RT G group were different from those in low RT G group. Only RT G revealed a difference between medium and low RT G groups. Correlation analysis showed that RT G was positively correlated with gender, age, BMI, HbA 1C, TC, and low density lipoprotein-cholesterol(LDL-C). Regression analysis showed that BMI, HbA 1C, and LDL-C were the related factors affecting the RT G of patients with type 2 diabetes. Conclusion:There is a larger proportion of patients with high RT G in type 2 diabetes mellitus. Their BMI, HbA 1C, and LDL-C are associated with RT G in the patients with type 2 diabetes mellitus.
7.Effect of SIRT1 gene silencing on radiosensitivity of diffuse large B-cell lymphoma cells
Yixin KANG ; Shegan GAO ; Yanzhen GUO ; Jun YAO ; Zhiye ZHANG ; Xiaohui GAO ; Dianbao ZHANG ; Shuangshuang GUO ; Lulin ZHANG
Chinese Journal of Radiation Oncology 2017;26(6):687-690
Objective To explore the effect of SIRT1 gene silencing on the radiosensitivity of diffuse large B-cell lymphoma (DLBCL) cells.Methods Immunohistochemistry was used to measure the protein expression of SIRT1 in DLBCL tissues.Western blot was used to measure the expression of SIRT1 in DLBCL cell lines (OCI-Ly3,SU-DHL-2,and SU-DHL-4) and the immortalized B cell line HMy2.CIR.After SU-DHL-4 cells were transfected with si-SIRT1 and si-NC using Lipofectamine 2000,the expression of SIRT1 was determined by Western blot.MTT assay and colony-forming assay were used to assess the cell growth and colony formation ability of SU-DHL-4 cells treated with radiation.The group t-test or univariate analysis of variance was used for comparison between groups.Results The expression rate of SIRT1 in DLBCL tissues was 72.6%(103/140),which was significantly higher than that in reactive lymphoid hyperplasia (RLH) tissues (26.5%,8/25)(P=0.001).The SIRT1 expression was significantly higher in DLBCL cells than in HMy2.CIR cells (P=0.020).After SIRT1 gene silencing by si-SIRT1,the expression of SIRT1 was significantly reduced in SU-DHL-4 cells (P=0.008).Besides,SIRT1 gene silencing significantly reduced the growth rate and colony formation ability of SU-DHL-4 cells treated with radiation (P=0.030).Conclusions SIRT1 gene silencing enhances the radiosensitivity of DLBCL cells,providing a novel target for the radiotherapy of DLBCL.
8.Measurement of corneal thickness by optical coherence tomography angiography
Peiwen ZHU ; Xuexiang ZOU ; Kangcheng LIU ; Yun HAN ; Zhirong LIN ; Lei YE ; Mei SHEN ; Honghua KANG ; Shuangshuang ZHOU ; Gang TAN ; Yi SHAO
Recent Advances in Ophthalmology 2017;37(8):732-735
Objective To analyze the thickness of cornea and corneal epithelium in healthy subjects by optical coherence tomography angiography (OCTA).Methods Totally 100 healthy subjects aged between 20 and 30 years were analyzed by OCTA technique.Using AngioVue OCTA system of retinal imaging mode,and using SSADA algorithm for imaging,the cornea and the corneal epithelium in the central corneal diameter range of 9 mm were measured.The differences of corneal and corneal epithelial thickness in different gender regions were compared.Results In the male and female group,the corneal central total thickness were (559.92 ±33.26) μm and(540.06 ±31.63)μm,and the corneal epithelial thickness were(57.78 ±4.88) μm and(56.88 ±4.57) μm,The total central corneal thickness and central corneal epithelial thickness of the male were greater than those of the female,the difference was statistically significant (t =3.06,2.10;all P < 0.05).The cornea of male was the thickest at S5,S7 and SN9,there were significant differences at S5 and S7 compared with female (t =2.93,2.83;all P < 0.05);The female cornea was the thickest at S5,SN7 and SN9,and the difference was significant at S5 compared with male.The cornea of male subjects was the thinnest at IT,which was statistically significant only at IT5 compared with female subjects in the same area (t =2.02,P < 0.05);The cornea of female subjects was the thinnest at T5,IT7 and T9,which was statistically significant only at T5 and T9 compared with male subjects in the same region (t =2.63,2.20;all P < 0.05);There was significant difference in corneal thickness between male and female at ST (t =3.1 1,2.79,2.33;all P < 0.05).The corneal epithelium was the thickest at IT5,I7,and I9,and the lowest at S5,S7 and S9,and there was no significant difference compared with female in the same region (all P > 0.05).The corneal epithelium of female at the IT5,T7,N9 were the thickest,SN5,S7,S9 were the thinnest;Except for M2 and SN5,there was no significant differences in corneal epithelium between male and female groups (all P > 0.05).Corneal central epithelium accounted for the largest percentage of total corneal thickness,and gradually decreased from inside to outside.Conclusion OCTA can be used to measure the thickness of corneal and corneal epithelial regions.
9.Effects of maternal position in the second stage of labor on delivery outcomes
Huixin ZHANG ; Rong HAO ; Hongyu ZHANG ; Xiaoli ZHANG ; Shuping ZHENG ; Huanhuan ZHANG ; Shuangshuang KANG
Chinese Journal of Perinatal Medicine 2014;17(11):743-747
Objective To evaluate the effects of the non-supine position and the supine position during the second stage of labor on delivery outcomes.Methods In total,642 singleton term primiparas in the Fourth Hospital of Hebei Medical University between January 2013 and March 2014 were divided into the non-supine position group (n=320),which included the lateral (n=204) and hands-and-knees position (n=116),and the supine position group (n=322) according to primipara's choice.Delivery outcomes in the two groups were compared using the Student's t test,Mann-Whitney rank sum test and the x2 test.Results The incidence of caesarean section was similar in the non-supine position and supine position groups [0.9%(3/320) vs 1.9% (6/322),x2=0.438,P=0.508].Compared with the supine position group,women in the non-supine position group had a higher incidence of perineal lacerations grede Ⅰ [85.2% (270/317) vs 65.2% (206/316),x2=33.884],longer duration of the second stage of labor [48.0 min (31.0-78.0 min) vs 41.0 min (25.0 67.3 min),Z=-3.066] and greater blood loss after 2 hours of labor [240.0 ml (200.0-287.5 ml) vs 210.0 ml (160.0-260.0 ml),Z=-3.736],but a significant reduction in posterolateral episiotomies [5.3%(17/317) vs 23.4% (74/316),x2=41.908],perinealedema [13.6% (43/317) vs 21.2% (67/316),x2=6.430] and meconiumstained liquor [20.8% (66/317) vs 33.2% (105/316),x2=12.356] (all P<0.05).In the non supine position group,fetal heart rate showed fewer early decelerations,variable decelerations and late decelerations than in the supine position group [10.1% (32/317) vs 17.1% (54/316),x2=6.593; 2.8% (9/317) vs 6.6% (21/316),x2=5.079; 3.2% (10/317) vs 7.6% (24/316),x2 6.139; all P<0.05,respectively].Conclusions The non supine position during the second stage of labor may improve delivery outcomes.

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