1.Proliferation-promoting effect of umbilical cord mesenchymal stem cells on co-cultured bovine mammary gland epithelial cells
Yankun ZHAO ; Wei SHAO ; Chenglong LUO ; Xiong YU
Acta Laboratorium Animalis Scientia Sinica 2017;25(4):391-398
Objective To explore the proliferation-promoting effect of bovine mammary gland epithelial cells (BMECs) co-cultured with umbilical cord mesenchymal stem cells (UC-MSCs) in serum-free culture mediuum.Methods Bovine UC-MSCs and BMECs were selected for co-culturing in direct or indirect contact.In the direct contact culture groups, UC-MSCs and BMECs were co-cultured at concentration ratios of 2∶1, 1∶1, 1∶2, 1∶3, 1∶4, 1∶5, and 1:10, respectively.In the indirect contact culture group, the supernatant of UC-MSCs was used as the conditioned medium to re-suspend BMECs.In the control groups, UC-MSCs and BMECs were cultured alone.The cell growth status in each group was observed at 0, 4, 8, 12, 24, 36, 48, 60, 72 h after culture, and cell proliferation was detected by cell counting kit-8 (CCK-8) assay.Results At 48 h, the optical density of the conditioned medium-BMECs group was significantly higher compared with the control groups (P<0.05).Meanwhile, the optical density in the direct contact group at a concentration ratio of 1∶2 reached the peak, which was extremely significantly higher compared with the control groups (P<0.01) and significantly higher compared with the other direct contact culture groups and the conditioned medium-BMECs group (P<0.05).Conclusions Co-culture of UC-MSCs and BMECs in serum-free culture medium is capable to promote the proliferation of BMECs, and the co-culture by cell-to-cell contact has a better effect.The optimal concentration ratio of UC-MSCs to BMECs is 1∶2, and the optimal culture time is 48 h.
2.Study on glomerular microvascular injury and repair in patients with IgA nephropathy and its relationship with intermedin
Junfeng LEI ; Yun ZHOU ; Honglin DONG ; Xiaoyu ZHAO ; Yankun LUO ; Rongshan LI
Chinese Journal of Nephrology 2017;33(6):416-421
Objective To investigate the glomerular microvascular injury and repair in patients with IgA nephropathy (IgAN) as well as its relationship with intermedin (IMD).Methods Eighty cases of renal tissue taken from patients first diagnosed as IgAN in Shanxi Provincial People's Hospital Affiliated to Shanxi Medical University and 15 cases of normal renal tissue were detected by the expression of glomerular IMD,CD31,and VE-cadherin through immunohistochemical method.ELISA method was used to detect VEGF and IMD of plasm from 31 normal subjects and 36 cases chosen from the IgAN patients.Their changes and internal relationship were analyzed according to Lee's and chronic kidney disease (CKD) classification.Results (1) Compared with the control group the expressions of CD31,IMD,and VE-cadherin in IgAN patients were statistically significant (P <0.01).Compared with the control group the levels of IMD and VEGF in plasma of IgAN patients in early stage of CKD group and late stage of CKD group were statistically significant (P < 0.01).(2)Correlation analysis:the expression of glomerular CD31 and Lee's classification were negatively correlated (r=-0.232,P < 0.05);glomerular IMD was negatively correlated with Lee's classification (r=-0.241,P<0.05),while positively correlated with glomerular VE-cadherin (r=0.417,P< 0.01).VEGF in plasma of IgAN patients was positive correlated with CKD classification,BUN (r=0.458,0.409,P<0.05),and negatively correlated with serum ALB (r=-0.532,P<0.01).Conclusion Microvascular injury exists in patients with IgAN.The expression of VE-cadherin and IMD are positively correlated,suggesting that IMD may be involved in the progression of vascular protection and angiogenesis in IgAN.The contents of IMD and VEGF in plasma of IgAN patients increase,indicating that they may play a role in the progression of IgAN.
3.Effect of calcium-sensing receptor on intracellular calcium, cell proliferation and migration of SGC-7901 cell line
Jian SUN ; Xiaoqin LIU ; Qi WU ; Li LI ; Hongtao ZHAO ; Yankun HAO ; Zhifang LANG ; Hairong LUAN
Chinese Journal of Clinical and Experimental Pathology 2015;(10):1140-1144
Purpose To observe the functional expression of calcium sensing receptor ( CaSR) in human gastric cancer SGC-7901 cell line, the effect of CaSR on intracellular calcium, cell proliferation and migration of SCG-7901. Methods The expression and distribu-tion of CaSR were detected by Western blotting and immunofluorescence observation in SGC-7901. The intracellular concentration of free calcium ( [ Ca2+] i ) was determined by confocal laser scanning microscopy. MTT, flow cytometry and scratch test were used to an-alyze the impact of CaSR the proliferation and the migration capabilities of SGC-7901 cell. Results CaSR protein was expressed in SGC-7901. Extracellular calcium or calindol significantly increased the expression of [Ca2+]i, CaSR and E-cadherin;In addition, the migration capabilities were decreased. Conclusion CaSR is expressed in SGC-7901. The activation of CaSR induces the expression of E-cadherin, and decreases migration ability.
4.Transcatheter closure treatment for patients with atrial septal defect complicated by atrial fibrillation:a postoperative follow-up study
Yankun YANG ; Hong ZHENG ; Zhengming XU ; Xin SUN ; Shiliang JIANG ; Zhongying XU ; Shihua ZHAO ; Gejun ZHANG ; Haibo HU ; Jinglin JIN ; Jianhua LV ; Tian LAN ; Fan YANG
Journal of Interventional Radiology 2014;(5):385-387
Objective To explore the prognosis and management of atrial fibrillation (AF) in patients with atrial septal defect(ASD) accompanied by AF after transcatheter closure of ASD. Methods During the period from July 2010 to May 2013, a total of 24 patients with ASD accompanied by AF were admitted to authors’ hospital to receive transcatheter closure of ASD. Electrocardiogram (ECG), chest X-ray film and transthoracic echocardiography (TTE) were performed before and one day after the operation. Follow-up information was obtained through telephone or at out-patient clinic interview. Results Successful occlusion of ASD was obtained in all patients, and in no patient the AF rhythm turned to sinus rhythm after the procedure. In one patient preoperative AF turned to postoperative atrial flutter, and AF recurred in one case who had received transcatheter ablation of AF before the procedure. One female patient developed gastric bleeding during the course of orally taking warfarin, and she died of cerebral infarction at three days after ceasing the use of warfarin. Of the 24 patients, no anticoagulant drug was used in 5 (20.8%), oral administration of aspirin was given in 7 (29.2%), and oral medication of warfarin was employed only in 11 (45.8%). Conclusion The spontaneous conversion rate of AF is very low in patients with ASD complicated by AF after transcatheter closure of ASD. Postoperative medication of anticoagulation should be strictly standardized and carefully managed.
5.Overexpression of miR-613 enhanced radiosensitivity of colorectal cancer cells via targeting downregulation of Wee1
Weishan ZHAO ; Yankun ZHU ; Ruotian WANG ; Aoran GUAN ; Ruhong LI
Journal of International Oncology 2019;46(3):157-165
Objective To explore the effect of microRNA-613 (miR-613)/Wee1 axis on the radiosensitivity of colorectal cancer cells.Methods A total of 20 patients with radiosensitive colorectal cancer and 20 patients with radioresistance were selected from Yan'an Hospital Affiliated to Kunming Medical University from November 2016 to May 2017.Human colorectal cancer cell lines LoVo and HCT116 were selected and the radioresistant cell lines LoVo/R and HCT116/R were established for subsequent experiments.Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of miR-613 and Wee1 in colorectal cancer tissues and cell lines.The radioresistant cells were transfected by miR-613 mimic,and non-transfected cells were used as control group.The effects of miR-613 overexpression on the proliferation,invasion and cell cycle of radiation resistance of colorectal cancer cells at different radiation doses were evaluated by CCK-8 assay,Transwell assay and Western blotting,respectively.Furthermore,dual-luciferase reporter gene assay was used to verify whether Wee1 was a target gene of miR-613.si-Wee1 was transfected into radioresistant cells of colorectal cancer,or co-transfected with si-Wee1 and miR-613 inhibitor,and non-transfected cells were used as control group.The effects of miR-613/Wee1 axis on cell proliferation,invasion and cell cycle were detected by CCK-8,Transwell and Western blotting at different radiation doses.Results The expression of miR-613 was downregulated in the radiation resistance group of patients (1.54 ± 0.25 vs.2.64 ± 0.45;t =3.140,P =0.009) and radiation resistance cell lines (LoVo/R vs.LoVo:1.03 ± 0.12 vs.3.05 ± 0.15;t =8.944,P =0.006;HCT116/R vs.HCT116:1.01 ±0.11 vs.2.85 ±0.16;t =8.050,P =0.008).Overexpression of miR-613 was significantly inhibited the proliferation (LoVo/R:t6 Gy =6.018,P =0.013;HCT116/R:t6Gy =5.634,P =0.015) and invasion (LoVo/R:45.00 ± 8.95 vs.180.15 ± 6.95,t6 Gy =11.93,P =0.003;HCT116/R:49.97 ±6.21 vs.170.20 ±7.03,t6 Gy =12.82,P =0.006) of LoVo/R and HCT116/R cells and decreased the expression levels of G2-M phase cell cycle correlated proteins (CDK1 and cyclin B).Moreover,dual-luciferase reporter gene assay confirmed that Wee1 was a target of miR-613.Mechanistically,overexpression of miR-613 promoted the radiosensitivity of LoVo/R and HCT116/R cells through inhibiting cell proliferation (compared with si-Wee1 group,co-transfected with si-Wee1 and miR-613 inhibitor,and control group,LoVo/R:F8 Gy =40.742,P =0.007;HCT116/R:F8 Gy =28.958,P =0.011),invasion (LoVo/R:F8 Gy =55.413,P =0.004;HCT116/R:F8 Gy =65.634,P =0.003) and arresting cell at G2-M phase via downregulating Wee1.Conclusion miR-613/Wee1 axis plays a certain role in regulating the radiation resistance of colorectal cancer cells,overexpression of miR-613 may reverse the radiation resistance of colorectal cancer cells.
6.Genotypic analysis of Mycobacterium tuberculosis strains collected from He'nan Province by 24-locus variable-number tandem-repeat typing and spoligotyping
Jie SHI ; Danwei ZHENG ; Yankun ZHU ; Junwei ZHAO ; Xiaoguang MA ; Shaohua WANG ; Hui LI ; Jin XING
Chinese Journal of Infectious Diseases 2018;36(12):741-746
Objective To investigate the genotype of M.tuberculosis in He'nan Province.Methods A total of 668 M.tuberculosis clinical strains collected in difference regions of He'nan Province during 2015 were genotyped by two standard methods,including classical 24-locus mycobacterium interspersed repetitive unit variable-number tandem-repeat (MIRU-VNTR) typing and spoligotyping.Results The 668 isolates were divided into 11 clusters and 35 patterns by spoligotyping.Among the 558 Beijing strains,546 were typical Beijing strains and the other 12 were atypical Beijing strains.Among the 110 non-Beijing strains,eight were new strains and the remaining 102 non-Beijing strains were divided into 10 families.There were 76 isolates belonging to T family,including 59 of T1 families,7 of T2 families,and 10 of T3 families.The 668 strains were divided into 550 gene patterns by standard 24-locus VNTR,including 508 un-clustered patterns and 160 clustered into 42 clusters.The largest cluster contained 21 strains,the other clusters contained 2-20 strains.Conclusion Beijing strain is still the most prevalent M.tuberculosis in He'nan Province.
7.Not Available.
Yaowen ZHANG ; Yankun LUO ; Jingqi ZHAO ; Wenzhuo ZHENG ; Jun ZHAN ; Huaping ZHENG ; Feng LUO
Acta Pharmaceutica Sinica B 2024;14(1):110-132
The aqueous two-phase system (ATPS) is an all-aqueous system fabricated from two immiscible aqueous phases. It is spontaneously assembled through physical liquid-liquid phase separation (LLPS) and can create suitable templates like the multicompartment of the intracellular environment. Delicate structures containing multiple compartments make it possible to endow materials with advanced functions. Due to the properties of ATPSs, ATPS-based drug delivery systems exhibit excellent biocompatibility, extraordinary loading efficiency, and intelligently controlled content release, which are particularly advantageous for delivering drugs in vivo. Therefore, we will systematically review and evaluate ATPSs as an ideal drug delivery system. Based on the basic mechanisms and influencing factors in forming ATPSs, the transformation of ATPSs into valuable biomaterials is described. Afterward, we concentrate on the most recent cutting-edge research on ATPS-based delivery systems. Finally, the potential for further collaborations between ATPS-based drug-carrying biomaterials and disease diagnosis and treatment is also explored.