1.Establishment of an in vitro model of brain blood barrier
Yunan YANG ; Yunhua GAO ; Jinning LOU ; Ping LIU ; Chixue FU
Journal of Third Military Medical University 2003;0(22):-
Objective To study the dynamic changes of transendothelial electrical resistance (TEER) on our in vitro model of brain blood barrier (BBB) made from primary culture of BALB/c mouse brain microvascular endothelial cells (BMVEC), and to explore the relation between TEER and BBB permeability, and search for the best culture condition. Methods BMVEC were isolated from BALB/c mouse and cultured on a transwell insert with special micro-pore. The cells were identified with immunohistochemical methods and electron microscope. TEER over BMVEC was measured after BBB model establishment for determining the 3H-Glucose permeability of BBB in vitro. Results BMVEC cultured in the transwell insert exhibited typical "flagstone" appearance and in a tight monolayer structure under electron microscope. Immunohistochemical detection of ZO-1 protein, a marker antigen of tight junction, showed smooth, continuous and tight junctions between confluent BMVEC. TEER over BMVEC monolayer increased to (346?10) ?/cm2 when the permeability for 3H-Glucose was decreased to the minimum. Conclusion BBB model in vitro made from primary culture of BMVEC in transwell has the basic characteristics of BBB in morphology, electrical resistance and permeability.
2.MR influence of different magnetic field and different scanner with same magnetic field on apparent diffusion coefficient of abdominal organs in healthy volunteers
Jiayin GAO ; Jiacheng ZHANG ; Zhenghan YANG ; Jinning LI ; Min CHEN ; Cheng ZHOU
Chinese Journal of Radiology 2013;(2):142-146
Objective To evaluate the reproducibility of ADC measurements at 1.5 vs 3.0 T and at 1.5 T of different scanners in liver,spleen and pancreas of healthy volunteers.Methods Abdominal DWI were performed on 33 healthy volunteers by using GE 1.5 T,Siemens 1.5 T and Philips 3.0 T MR scanners.The mean ADC values of liver,spleen,pancreatic head,body,and tail were calculated.The ADC data were analyzed by using paired-sample t tests.Results The mean ADC of liver at GE 1.5 T,Siemens 1.5T and Philips 3.0 T were (1.56 ±0.10) ×10-3,(1.67 ±0.15) ×10-3 and(1.35 ±0.12) ×10-3 mm2/s,spleen were (0.96±0.10) × 10 3,(0.98 ±0.11) ×10-3and(0.81 ±0.14) × 10-3 mm2/s,pancreatic head were (2.09 ± 0.27) × 10-3,(2.20 ± 0.21) × 10-3 and (2.05 ± 0.27) × 10-3 mm2/s,pancreatic body were (2.03 ± 0.27) × 10-3,(2.09 ± 0.30) × 10-3 and (1.76 ± 0.25) × 10-3 mm2/s,pancreatic tail were (1.88 ± 0.28) × 10-3,(1.88 ± 0.27) × 10-3 and (1.56 ± 0.27) × 10-3 mm2/s,respectively.From the aspect of different field strength MR scanners,there were significant differences in mean ADC of liver (t =11.073,P <0.01 in GE 1.5 T vs Philips 3.0 T; t =12.795,P <0.01 in Siemens 1.5 T vs Philips 3.0 T),spleen (t =4.143,P < 0.01 in GE 1.5 T vs Philips 3.0 T; t =5.376,P < 0.01 in Siemens 1.5 T vs Philips 3.0 T),pancreatic body (t =4.677,P < 0.01 in GE 1.5 T vs Philips 3.0 T; t =5.174,P <0.01 in Siemens 1.5 T vs Philips 3.0 T) and tail (t =5.356,P <0.01 in GE 1.5 T vs Philips 3.0 T; t =4.648,P <0.01 in Siemens 1.5 T vs Philips 3.0 T),but there were no significant differences in mean ADC of pancreatic head (t =0.340,P > 0.05 in GE 1.5 T vs Philips 3.0 T; t =1.349,P > 0.05 in Siemens 1.5 T vs Philips3.0 T).From the aspect of different 1.5 T MR scanners,there were significant differences in mean ADC of liver (t =-4.563,P < 0.01),but there were no significant differences in mean ADC of spleen (t =-0.732,P > 0.05),pancreatic head (t =-0.879,P > 0.05),body (t =-1.020,P >0.05) and tail (t =0.054,P > 0.05).Conclusion Between 1.5 T and 3.0 T MR scanners,there were significant differences in mean ADC of liver,spleen,pancreatic body and tail,but there were no significant differences in mean ADC of pancreatic head.At different 1.5 T MR scanners,there were significant differences in mean ADC of liver,but there were no significant differences in mean ADC of spleen,pancreatic head,body and tail.
3.Construction and evaluation of diabetic atherosclerosis model in LDLR-/-mice in-duced by STZ and fed with high-fat diet
Li WANG ; Jinning GAO ; Xiangyun CHEN ; Peiqing JIANG ; Mei LI ; Fangfang BU ; Mei ZHANG ; Guihua YAO ; Hui SUN
Chinese Journal of Arteriosclerosis 2024;32(11):985-993
Aim To construct a diabetic atherosclerosis mouse model and study the pathological characteristics of diabetic atherosclerosis.Methods Fifty 8-week-old male LDLR-/-mice were fed with standard diet for 2 weeks and then changed to high-fat diet,they were randomly divided into two groups.The diabetic atherosclerosis group was given intraperitoneal injection of low dose streptozotocin(STZ)for 5 days continuouly to establish the model,and the atheroscle-rosis group was given citrate buffer injection at the same time.The body mass,blood glucose and blood lipids of the mice in the two groups were detected for many times.At the age of 23 weeks,the mice were euthanized after glucose tolerance test.HE staining and oil red O staining were used to detect the gross and aortic root atherosclerosis,immunohistochemical staining was used to detect CD4,α-smooth muscle actin(α-SMA),EGF-like module-containing mucin-like hormone re-ceptor-like 1(EMR1),monocyte chemotactic protein-1(MCP-1),NOD-like receptor protein 3(NLRP3),vascular cell adhesion molecule-1(VCAM-1),matrix metalloproteinase-2(MMP-2)and tissue inhibitor of metalloproteinase-1(TIMP-1),Western blot was used to detect α-SMA,CD4,tumor necrosis factor-α(TNF-α),NLPR3,intercellular adhesion molecule-1(ICAM-1),and type Ⅰ and Ⅲ collagen.Results Compared with the atherosclerosis group,the body mass decreased,the levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDLC)increased,and the levels of high density lipoprotein cholesterol(HDLC)decreased(P<0.05)in the diabetic atherosclerosis group.Compared with the atherosclerosis group,the distribution of atherosclerotic plaques was diffuse and the area was increased in the diabetic atherosclerosis group,and the contents of lipids,T cells,macrophages,smooth muscle cells,type Ⅰ and Ⅲ colla-gen were increased(P<0.05);the protein levels of TNF-α,MCP-1,MMP-2,NLRP3,ICAM-1 and VCAM-1 in vascular tissues were increased,while the content of TIMP-1 were decreased and MMP2/TIMP-1 were increased(P<0.05).Conclusions LDLR-mouse model of diabetic atherosclerosis can be successfully established by STZ induction combined with high-fat diet,which can reflect the plaque composition and inflammatory characteristics of diabetes promoting atheroscle-rosis.It can be used as a relatively ideal pathological model for the study of diabetic macroangiopathy.
4.Single-cell Sequencing Reveals Clearance of Blastula Chromosomal Mosaicism in In Vitro Fertilization Babies
Gao YUAN ; Zhang JINNING ; Liu ZHENYU ; Qi SHUYUE ; Guo XINMENG ; Wang HUI ; Cheng YANFEI ; Tian SHUANG ; Ma MINYUE ; Peng HONGMEI ; Wen LU ; Tang FUCHOU ; Yao YUANQING
Genomics, Proteomics & Bioinformatics 2022;(6):1224-1231
Although chromosomal mosaic embryos detected by trophectoderm(TE)biopsy offer healthy embryos available for transfer,high-resolution postnatal karyotyping and chromosome testing of the transferred embryos are insufficient.Here,we applied single-cell multi-omics sequenc-ing for seven infants with blastula chromosomal mosaicism detected by TE biopsy.The chromo-some ploidy was examined by single-cell genome analysis,with the cellular identity being identified by single-cell transcriptome analysis.A total of 1616 peripheral leukocytes from seven infants with embryonic chromosomal mosaicism and three control ones with euploid TE biopsy were analyzed.A small number of blood cells showed copy number alterations(CNAs)on seem-ingly random locations at a frequency of 0%-2.5%per infant.However,none of the cells showed CNAs that were the same as those of the corresponding TE biopsies.The blastula chromosomal mosaicism may be fully self-corrected,probably through the selective loss of the aneuploid cells dur-ing development,and the transferred embryos can be born as euploid infants without mosaic CNAs corresponding to the TE biopsies.The results provide a new reference for the evaluations of trans-ferring chromosomal mosaic embryos in certain situations.
5. Efficacy of femoral triangle versus adductor canal approach to saphenous nerve block for postoperative analgesia in patients undergoing knee arthroplasty
Chunguang WANG ; Zhiqiang ZHANG ; Yanjun LI ; Yanhui BAI ; Yuanyuan WANG ; Qinghui LI ; Jiayun LIU ; Jinning LIU ; Meina GAO ; Xiaoyu GUO
Chinese Journal of Anesthesiology 2019;39(8):953-956
Objective:
To compare the efficacy of femoral triangle versus adductor canal approach to saphenous nerve block for postoperative analgesia in the patients undergoing knee arthroplasty.
Methods:
Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients of both sexes, aged 53-68 yr, scheduled for elective total knee arthroplasty under general anesthesia, were assigned into 2 groups (