1.Experimental study on homeobox B8 in promoting metastasis of HGSOC by regulating H3K27me3 modification of KDM6B-mediated C/EBPα histone
Li XIANG ; Donghua WANG ; Ping WANG ; Shixiong GONG ; Yajun HU
China Medical Equipment 2025;22(11):164-173
Objective:To research the mechanism of the regulation of homeobox B8(HOXB8)for lysine demethylase 6B(KDM6B)-mediated CCAAT/enhancer binding protein α(C/EBPα)axis on the metastasis of high-grade serous ovarian cancer(HGSOC),so as to provide references for the study of the pathogenesis of HGSOC patients.Methods:The tumor tissue samples and corresponding adjacent normal tissue samples of HGSOC patients admitted to Wuhan Hospital of Traditional Chinese and Western Medicine from June to December 2024 were selected,and cell lines SKOV3 and A2780 of ovarian cancer were collected.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was adopted to detect the mRNA levels of KDM6B in HGSOC tumor tissues and its corresponding adjacent tissues.Western blot assay and immunohistochemistry were adopted to detect the expressions of KDM6B protein in the tissue.The A2780 cells of ovarian cancer were divided into the oe-HOXB8 group that was transfected by the HOXB8 overexpression vector,and the oe-NCHOXB8 group with the negative control(NC)vector.The SKOV3 cells of ovarian cancer were divided into the si-HOXB8 group that was transfected by the HOXB8 small interference sequence,and the si-NCHOXB8 group with negative control sequence.The transfected KDM6B was divided into the si-KDM6B group with small interference sequence and the oe-KDM6B group transfected with overexpression vector.The co-transfection HOXB8 and(or)KDM6B,C/EBPα were divided into si-HOXB8+si-KDM6B group and si-HOXB8+si-C/EBPα group of small interference sequence.The chromatin immunoprecipitation-qPCR(ChIP-qPCR)and dual-luciferase reporter gene assay were used to verify the mechanism that HOXB8 transcript and regulate KDM6B in SKOV3 and A2780 cells of ovarian cancer.The effects of overexpression or silencing of HOXB8 in A2780 and SKOV3 cells on the proliferation,invasion,migration and KDM6B expression of ovarian cancer cells were detected.The effects of overexpression or silencing of KDM6B in SKOV3 cells on the trimethylation modification of lysine 27 at histone H3(H3K27me3)and the expression of C/EBPα were detected.The effects of silencing KDM6B and C/EBPα on HOXB8-induced cell proliferation,invasion and migration were analyzed through functional rescue experiments.Results:In tumor tissues,the mRNA and protein expression levels of KDM6B were 1.02±0.03 and 1.02±0.04,respectively,which were significantly higher than those in the adjacent tissues,and the differences were statistically significant(t=62.440,38.737,P<0.01).The optical density value of proliferation,invasion rate and migration rate of A2780 cells in the oe-HOXB8 group that was transfected by the HOXB8 overexpression vector were respectively(1.74±0.15),(89.71±6.60)%and(85.33%±7.02)%,which were significantly higher than those in the oe-NCHOXB8 group,and the differences were statistically significant(t=7.778,7.353,4.759,P<0.01).The optical density value of proliferation,invasion rate and migration rate of SKOV3 cells in the si-HOXB8 group were respectively(0.54±0.06),(47.23±3.41)%and(43.20±3.12)%,all of which were significantly lower than those in the si-NCHOXB8 group,and the differences were statistically significant(t=9.400,8.615,9.040,P<0.01).The optical density value of proliferation,invasion rate and migration rate of SKOV3 cells in the si-HOXB8+si-KDM6B group were(1.04±0.09),(73.11±4.98)%and(68.65±4.45)%,respectively,which were significantly higher than those in the si-HOXB8 group,and the differences were all statistically significant(t=6.875,6.852,7.562,P<0.01).The optical density value of proliferation,invasion rate and migration rate of SKOV3 cells in the si-HOXB8+si-C/EBPα group were respectively(0.97±0.07),(75.87±5.12)%and(70.59±4.81)%,all of which were significantly higher than those in the si-HOXB8 group,and the differences were all statistically significant(t=6.355,7.500,7.884,P<0.01).Conclusion:HOXB8 can inhibit the C/EBPα expression and promote the HGSOC metastasis by regulating and controlling H3K27me3 modification of KDM6B-mediated C/EBPα histone.
2.Experimental study on homeobox B8 in promoting metastasis of HGSOC by regulating H3K27me3 modification of KDM6B-mediated C/EBPα histone
Li XIANG ; Donghua WANG ; Ping WANG ; Shixiong GONG ; Yajun HU
China Medical Equipment 2025;22(11):164-173
Objective:To research the mechanism of the regulation of homeobox B8(HOXB8)for lysine demethylase 6B(KDM6B)-mediated CCAAT/enhancer binding protein α(C/EBPα)axis on the metastasis of high-grade serous ovarian cancer(HGSOC),so as to provide references for the study of the pathogenesis of HGSOC patients.Methods:The tumor tissue samples and corresponding adjacent normal tissue samples of HGSOC patients admitted to Wuhan Hospital of Traditional Chinese and Western Medicine from June to December 2024 were selected,and cell lines SKOV3 and A2780 of ovarian cancer were collected.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was adopted to detect the mRNA levels of KDM6B in HGSOC tumor tissues and its corresponding adjacent tissues.Western blot assay and immunohistochemistry were adopted to detect the expressions of KDM6B protein in the tissue.The A2780 cells of ovarian cancer were divided into the oe-HOXB8 group that was transfected by the HOXB8 overexpression vector,and the oe-NCHOXB8 group with the negative control(NC)vector.The SKOV3 cells of ovarian cancer were divided into the si-HOXB8 group that was transfected by the HOXB8 small interference sequence,and the si-NCHOXB8 group with negative control sequence.The transfected KDM6B was divided into the si-KDM6B group with small interference sequence and the oe-KDM6B group transfected with overexpression vector.The co-transfection HOXB8 and(or)KDM6B,C/EBPα were divided into si-HOXB8+si-KDM6B group and si-HOXB8+si-C/EBPα group of small interference sequence.The chromatin immunoprecipitation-qPCR(ChIP-qPCR)and dual-luciferase reporter gene assay were used to verify the mechanism that HOXB8 transcript and regulate KDM6B in SKOV3 and A2780 cells of ovarian cancer.The effects of overexpression or silencing of HOXB8 in A2780 and SKOV3 cells on the proliferation,invasion,migration and KDM6B expression of ovarian cancer cells were detected.The effects of overexpression or silencing of KDM6B in SKOV3 cells on the trimethylation modification of lysine 27 at histone H3(H3K27me3)and the expression of C/EBPα were detected.The effects of silencing KDM6B and C/EBPα on HOXB8-induced cell proliferation,invasion and migration were analyzed through functional rescue experiments.Results:In tumor tissues,the mRNA and protein expression levels of KDM6B were 1.02±0.03 and 1.02±0.04,respectively,which were significantly higher than those in the adjacent tissues,and the differences were statistically significant(t=62.440,38.737,P<0.01).The optical density value of proliferation,invasion rate and migration rate of A2780 cells in the oe-HOXB8 group that was transfected by the HOXB8 overexpression vector were respectively(1.74±0.15),(89.71±6.60)%and(85.33%±7.02)%,which were significantly higher than those in the oe-NCHOXB8 group,and the differences were statistically significant(t=7.778,7.353,4.759,P<0.01).The optical density value of proliferation,invasion rate and migration rate of SKOV3 cells in the si-HOXB8 group were respectively(0.54±0.06),(47.23±3.41)%and(43.20±3.12)%,all of which were significantly lower than those in the si-NCHOXB8 group,and the differences were statistically significant(t=9.400,8.615,9.040,P<0.01).The optical density value of proliferation,invasion rate and migration rate of SKOV3 cells in the si-HOXB8+si-KDM6B group were(1.04±0.09),(73.11±4.98)%and(68.65±4.45)%,respectively,which were significantly higher than those in the si-HOXB8 group,and the differences were all statistically significant(t=6.875,6.852,7.562,P<0.01).The optical density value of proliferation,invasion rate and migration rate of SKOV3 cells in the si-HOXB8+si-C/EBPα group were respectively(0.97±0.07),(75.87±5.12)%and(70.59±4.81)%,all of which were significantly higher than those in the si-HOXB8 group,and the differences were all statistically significant(t=6.355,7.500,7.884,P<0.01).Conclusion:HOXB8 can inhibit the C/EBPα expression and promote the HGSOC metastasis by regulating and controlling H3K27me3 modification of KDM6B-mediated C/EBPα histone.
3.The application of laparoscopic curettage in the standardized training for residents
Zhongxian WANG ; Wei LÜ ; Li XIANG ; Shixiong GONG ; Yi HE
Chinese Journal of Medical Education Research 2019;18(5):519-522
Objective To study the application value of laparoscopic curettage in the standardized training for residents.Methods In this study,we selected residents of the Wuhan First Hospital as the research subjects.Thirty of the subjects from the 2015 grade were the control group and another 29 from the 2016 grade were the experimental group.Both groups had three classes,including one theory class,and two practice classes.After the theoretical class,the experimental group practiced laparoscopic curettage,while the control group practiced curettage.After practice,the experimental group filled out the questionnaire on teaching quality,and both groups participated in the theory test.Results Laparoscopic curettage can significantly improve students' ability to learn and understand curettage,so that most students can adapt to curettage,eliminate the panic from uterine perforation and be more sensitive to the uterus textures,and satisfactory teaching results were achieved.The average score of the practice test for the experimental group (9.150 ± 2.461) was statistically higher than that for the control group (7.053 ± 1.347).The average score of the theory test (7.581 ± 1.632) for the experimental group was not statistically different from that for the control group (6.983 ± 1.726).The average scores of the two tests for the experimental group (9.037 ± 1.951) and the control group (6.891 ± 2.140) were statistically different.Conclusions Laparoscopic curettage can significantly improve the ability to learn,understand and feel curettage for residents who are under standardized training,and it has great application value in the three basic operation trainings.
4.Clinical and epidemic characteristics of 27 cases with Brucellosis confirmed by bacterial culture
Kaizhong LUO ; Wei CAO ; Min WANG ; Shixiong HU ; Shaojun ZHANG ; Guozhong GONG
Journal of Chinese Physician 2017;19(11):1654-1657
Objective To improve the diagnostic and therapeutic ability of Brucellosis by analyzing the epidemic and clinical characteristics.Methods A retrospective analysis was done on the data of Brucellosis patients treated in our hospital from 2007 to 2016.Results Since the first case was diagnosed in 2012,27 patients [19 male and 8 female,mean age (44.4 ± 16.9) years] were confirmed by clinical manifestations and positive bacterial cultures results.The annual number of cases from 2012 to 2016 was 1,1,6,4 and 15.Among them,10 cases (37.0%) had a history of close contact with goat,7 cases (25.9%)with raw mutton,1 case (3.7%) with raw beef and 1 case (3.7%) with suspicious laboratory contamination while 8 cases (29.6%) had no evident risk factors for Brucellosis.The common clinical manifestation included fever (81.5%),lumbago/joint pain (55.6%),fatigue (33.3%) and hyperhidrosis (22.2%).The white blood cell count was normal among 20 cases (74.1%) while 6 cases (22.2%) with leukopenia.Mild to moderate anemia in 20 cases (74.1%) and decreased platelet number in 4 case (14.8%).The percentage of elevated alanine aminotransferase,aspartate aminotransferase and lactate dehydrogenase was 32.0%,48.0% and 100%,respectively.Decreased albumin level was found in 23 cases (92.0%).The percentage of elevated erythrocyte sedimentation rate (ESR),C-reactive protein and serum ferritin 75.0%,82.3% and 77.8%,respectively,while 12 cases (85.7%) with procalcitonin level below 0.5 ng/L.According to follow-up for at least half year,all the cases were cured by active medical management.Conclusions The number of Brucellosis cases is rapidly increasing in our hospital.It's of great significance to know the epidemic and clinical characteristics of Brucellosis.

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