KDM4A promotes the migration and invasion of breast cancer cell line MDA-MB-231 by downregulating BMP9 / 中国癌症杂志

Background and purpose:Exogenous bone morphogenetic protein 9(BMP9)inhibits the malignant progression of human breast cancer,but its expression is often abnormally low in breast cancer.In this study,we intended to explore the expression and role of epigenetically-modified histone lysine-specific demethylase 4A(KDM4A)in breast cancer,and to investigate the relationship between KDM4A and BMP9 and its possible regulatory mechanism.Methods:The expression of KDM4A in breast cancer and its relationship with BMP9 were analyzed by bioinformatics and verified by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.Chromatin immunoprecipitation(ChIP)verified the regulatory role of KDM4A on BMP9,and RNA stability experiments and CHX protein stability experiments verified the effect of KDM4A in BMP9 expression.Exogenous recombinant MDA-MB-231 cells transfected with KDM4A small interfering RNA(siKDM4A)or infected with siBMP9 adenovirus(Ad-siBMP9)were constructed using RNA interference technology and adenoviruses knocking down BMP9,and the migratory and invasive abilities of the cells were detected by scratch healing assay and transwell assay,respectively.Results:Bioinformatics analysis showed that the expression of KDM4A was significantly higher in breast cancer than in normal tissues,and there was a negative correlation between the expression of KDM4A and that of BMP9 in breast cancer;RTFQ-PCR and Western blot showed that KDM4A was highly expressed in different breast cancer cell lines,and the knockdown of KDM4A significantly up-regulated BMP9.ChIP experiment confirmed that KDM4A could be significantly enriched in the promoter region of BMP9 gene,reducing its histone lysine 36 position instead of position 4 methyl status,thus silencing the expression of BMP9.RNA stability assay and CHX protein stability assay confirmed that KDM4A had no significant effect on the mRNA of BMP9,but could affect its protein degradation.After knocking down KDM4A,the migration and invasion abilities of breast cancer cells MDA-MB-231 were significantly inhibited,and this effect could be partially reversed by knocking down BMP9.Conclusion:KDM4A is highly expressed in breast cancer and breast cancer cell MDA-MB-231,and can silence its expression by down-regulating the level of histone methylation in the promoter region of the BMP9 gene,as well as affecting the stability of BMP9 at the protein level rather than at the level of mRNA,and promoting the migration and invasion of breast cancer.

Clinical application and efficacy analysis of castor stent graft in the treatment of aortic diseases / 中华胸心血管外科杂志

Objective:To summarize the use of Castor stent graft in aortic diseases and to analyze their efficacy.Methods:The clinical data of patients with aortic diseases treated with Castor stent graft from November 2017 to August 2022 in Fujian Provincial Hospital were collected and divided into branched stent group and branched stent co-operative group according to the operation method, and the clinical data of both groups were summarized.Results:A total of 75 cases of aortic disease were treated with Castor stents, and finally 53 cases were classified as branched stent group and 22 cases as branched stent co-operative group. The operations in both groups were successful. The median operative time in the branched stent group was 120(100, 160)min, and the median postoperative hospital stay was 7.0(5.5, 10.5)days.There was no postoperative ischemic stroke, no spinal cord ischemia. One case of new aortic dissection occurred. During follow-up, there was one lost case and two deaths, and the rest did not have endoleak, branch stent stenosis, ischemic stroke or re-intervention. In the branched stent co-operative group, there was one postoperative ischemic stroke, one case of slight stenosis of the left subclavian artery stent during follow-up, the remaining cases had satisfactory postoperative outcomes.Conclusion:Castor stent graft is a safe and effective procedure in the treatment of aortic diseases. And Castor stent graft can be used in combination with other endovascular repair techniques in the treatment of complex aortic diseases, with safe and reliable postoperative outcomes.

Application of fenestration technique in the treatment of aortic arch disease / 中华胸心血管外科杂志

Objective Assessment of fenestration in the aortic arch disease .Methods 13 patients with aortic arch dis-ease underwent fenestration operation to reconstruct affiliated vessels , aortic arch aneurysm in 5 patients, aortic anch ulcer in 4, Ⅲ type aortic dissection in 3 and Ⅰ type leakage after aortic stent graft in 1 patient.All patieuts were divided into 2 groups.8 patients in pre-fenestration group, 5 patients in in-situ fenestration group.Only left subclavian artery was rebuilded in 9 patients, both left carotid common artery and left subclavian artery were rebuilded in 3 patients, all three affiliated vessels of arch were rebuilded in 1 patient.Results Branch vessels of aortic arch were successfully rebuilded in all 13 patients.There were no endoleak, stenosis of branch vessels, graft diaplacement or deaths peri-operative period.During follow-up, no postop-erative complications occurred and all target vessels remained patent .No fenestration related endoleaks were observed.Conclu-sion Fenestration may be a viable alternative for patients with aortic arch disease .

Hybrid aortic arch replacement for aortic arch disease / 中华胸心血管外科杂志

Objective we assessed our institutional outcomes of hybrid treatment for aortic arch disease with supra-aortic debranching and endovascular stent graft repair.Methods From March 2016 to November 2016,6 patients underwent Hybrid total aortic arch repair:1 had aortic arch pseudoaneurysm;1 had type Ⅲ aortic dissection;4 had aortic arch aneurysm because of hypotension,of whom 1 with aneurysm prerupture and 1 with Aortic intramural hematoma.Supra-aortic vessels were involved and high-risk for traditional operations in all patients.Bifurcated artificial vessels were used,main vessel was end-to-side anastomosed with ascending aorta.Branching vessel were end-to-end anastomosed with right innominate artery and left subclavian artery,end-to-side anastomosed with left common carotid artery.Then,stent graft was implanted into ascending aorta and aortic arch.All patients were followed postoperatively,with regularly contrast computed tomography angiogram (CTA) and echocardiography(discharge,three months,six months,and yearly).Results Hybrid procedure with supra-aortic debranching and endovascular stent graft repair were completed in all patients,technical success rate was 100％.There were no perioperative obvious morbidity and mortality,follow-up period were 2-9 months.1 patients had stroke during follow-up period,condition improved after treatment.Supra-aortic vessels were patency and there were no endoleak in all patients.There were no recurrent aortic disease during follow-up period.Conclusion Hybrid aortic arch replacement can be performed with good postoperative and early results in high-risk patients for traditional open repair.

Construction of prokaryotic expression vector of HPV16E6 gene and its expression / 生物医学工程学杂志

Prokaryotic expression vector of mouse HPV16E6 gene was constructed. A pair of primers were designed according to the digestion sites in plasmid pGEX-KG and the HPV16E6 gene sequence published by GenBank. The DNA fragment of 321bp was amplified by PCR from the HPV recombinant plasmid with HPV16E6 gene, then cloned into pGEX-KG and transformed into the host E. coli strain JM109. The fragment was conformed to the original sequence, which indicated that fusion expression vector pGEX-KG-HPV16E6 was constructed. The pGEX-KG-HPV16E6 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degrees C, the expression product of HPV16E6 gene was identified by SDS-PAGE and Western blot. HPV16E6 fusion protein had been expressed successfully in the form of inclusion bodies, the molecular weight of fusion protein being 38 kD. Meanwhile, the optimum condition of HPV16E6 fusion protein expression was induced with 1.0 mmol/L IPTG for 4h. The fusion protein reacted specifically with the antibodies against HPV16E6. HPV16E6 gene was successfully expressed in E. coli, which could be used as a basis for preparing HPV16E6 vaccine in human.

Cloning and Expression of HPV18 E6 Gene / 天津医药

Objective: To express the protein of HPV18E6 based on pET-32a(+) at high level and study the expression and significance of HPV18E6 proteins in laryngeal carcinoma. Methods: The HPV18E6 gene was amplified by PCR and cloned into pET-32a(+). The amplified fragment was inserted into the plasmid pET32a (+) that was digested with BamHⅠand Hind Ⅲ. The recombinant plasmid pET32/E6 was transformed into E.coli JM109 which was selected with ampicillin. The recombinant plasmids were successfully introduced into E.coli BL21(DE 3) and were induced by IPTG. SDS-PAGE and Western blot analysis were used to detect the confusion protein. Finally, the optimization of expression conditions, such as temperature, concentration of IPTG, was studied. Results: The recombinant plasmids were identified and confirmed with enzyme digestion and sequencing. The BL21(DE3) transformed recombinant plasmid pET32/E6 had expressed HPV18E6 recombinant protein effectively. The optimum conditions of expression were 37 ℃, 1 mmol/L IPTG. Conclusion:Prokaryotic expression vector pET-32a(+)-HPV18E6 was successfully constructed. The high-level expression of HPV18E6 was achieved in E.coli BL21(DE3).