3.Effects of Col-Tgel, a bioactive material, on human tissue stem cells
Xiu-Xiu YIN ; Lin-Ping HU ; Cai-Ying ZHU ; Xiao-Bing ZHANG ; Tao CHENG
Chinese Journal of Tissue Engineering Research 2018;22(10):1540-1546
BACKGROUND:The traditional two-dimensional culture system has been widely used in the in vitro culture of human tissue stem cells,but it cannot really simulate the three-dimensional physiological microenvironment in the body, which is not conducive to the study of the biological behavior of human stem cells. OBJECTIVE: To detect the effect of the bioactivity of Col-Tgel in human hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs)in vitro and in vivo,by constructing a three-dimensional culture system stimulating the physiological microenvironment of the body. METHODS:(1)In vitro co-culture:Green fluorescent protein labeled MSCs(MSCs-GFP)and human umbilical cord blood CD34+cells were co-cultured in Col-Tgel for 3 days (three-dimensional culture group). Human umbilical cord blood CD34+cells were cultured in Col-Tgel for 3 days as single culture group. MSCs-GFP and human umbilical cord blood CD34+cells were co-cultured in Transwell chamber for 3 days as two-dimensional culture group. Human umbilical cord blood CD34+cells were cultured routinely as control group. The percentage of CD34+CD38-CD45RA-CD90+cells in each group was measured by flow cytometry. In situ immunofluorescence staining was used to detect the activity of cells that were co-cultured in Col-Tgel.(2)In vivo transplantation:NOD/SCID mice subjected to 24-hour X-ray irradiation were divided into two groups: in experimental group, MSC-GFP cells were resuspended in Col-Tgel and transplanted into the tibia of NOD/SCID mice; in control group, MSCs-GFP were resuspended in PBS and transplanted into the tibia of NOD/SCID mice. The MSC-GFP growth in the bone marrow was detected by two-photon/confocal microscopy at 3 days post transplantation. RESULTS AND CONCLUSION: (1) After co-culture in Col-Tgel for 3 days, the percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was 2.8 times that of the two-dimensional culture group, indicating that the MSCs significantly promoted the expansion of CD34+CD38-CD45RA-CD90+cells in the Col-Tgel. The percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was increased by 4.5 times compared with the single culture group and increased by 1.5 times compared with the control group. Immunofluorescence staining showed that the cell viability of human MSCs and human umbilical cord blood CD34+cells was not affected after co-cultured in Col-Tgel for 3 days.In the in vivo transplantation experiment,MSC-GFP cells could survive in the medullary cavity.In summary, Col-Tgel provides a new strategy for stem cell culture and in vivo growth by forming a three-dimensional system similar to the physiological environment in vivo.
4.Application of high-resolution ultrasound and CDFI-guided minimally invasive operation for breast lesions
De-Zhan WEI ; Xiu-Ying LU ; Da-Jiang ZHU ; Yi-Ye DENG ; Hong-Xia LI ; Xiu-Zhen DENG ; Zhan-Kui LU ; Chao CHENG ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(07):-
Objective To evaluate the application and the good qualities of high-resolution ultrasound and CDFI-guided mammotome minimally invasive biopsy device in the diagnosis and treatment of breast lesions.Methods The common clinical operations and the lesions which were guided mammotome minimally invasive biopsy device by high-resolution ultrasound and CDFI were contrasted.The effects of treatment were evaluated.Results 307 le- sions of 102 patients were removed by this method,and the operational process was successful.Patients' skin lacera- tions were tiny.Only one lesion was clinically diagnosed as mild blood clot under skin,but without other complica- tions.Conclusion Contrasted with the common clinily operations.the high-resolution ultrasound and CDFI-guided mammotome minimally invasive biopsy device in the diagnosis and treatment of breast lesion is effective,and the scar is tiny.It releases patients' pain.
5.Clinicopathologic study of aortic valves in children.
Ping HUANG ; Hong-wei WANG ; Zhen-lu ZHANG ; Xiu-fen HU ; Yan-ping LI ; Pei-xuan CHENG ; Jian-ying LIU
Chinese Journal of Pathology 2006;35(10):623-624
Adolescent
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Aortic Valve
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abnormalities
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Aortic Valve Insufficiency
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complications
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pathology
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surgery
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Aortic Valve Stenosis
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complications
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pathology
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surgery
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Child
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Endocarditis
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complications
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pathology
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surgery
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Female
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Heart Defects, Congenital
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complications
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pathology
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surgery
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Heart Valve Prosthesis Implantation
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Humans
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Male
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Rheumatic Heart Disease
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complications
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pathology
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surgery
6.Clinical study on the stability of palatal implant anchorage.
Ying ZHAO ; Yu-cheng SU ; Xiu-ying JIANG ; Juan DU
Chinese Journal of Stomatology 2005;40(6):463-467
OBJECTIVETo evaluate the clinical stability of the palatal implant anchorage system in orthodontic treatment.
METHODSSixteen osseointegrated implants (5.0 mm in diameter, 6 mm length) were inserted in the median palatal suture area of 19 patients with malocclusion (average age: 18.22 +/- 7.10 years, from 11 years to 35 years 1 month) as anchorage of active orthodontic treatment with MBT appliance. The standard lateral cephalogram after implant placement and before implant removing was taken to compare the radiological parameters.
RESULTSThe successful rate of palatal implant anchorage was 84.2%. The average duration of 16 palatal implant was 23.08 +/- 8.06 months (from 10 months to 36 months). There were no statistical differences in all parameters from the implant placement to the end of treatment. IL-X was from (62.88 +/- 5.85) mm to (62.45 +/- 6.70) mm, IL-Y was from (36.66 +/- 5.41) mm to (37.96 +/- 4.90) mm, IAP-PP was from (73.81 +/- 8.84) degrees to (74.72 +/- 9.22) degrees, IAP-Y was from (62.09 +/- 9.33) degrees to (63.85 +/- 10.96) degrees, U6-Y is from (20.80 +/- 5.87) mm to (21.49 +/- 6.00) mm.
CONCLUSIONSStable palatal implant anchorage was maintained during active orthodontic treatment.
Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Malocclusion ; therapy ; Orthodontic Anchorage Procedures ; instrumentation ; Orthodontic Appliance Design ; Palate, Hard ; surgery ; Young Adult
7.Establishment of a high expressing system of human coagulant factor VIII in vitro.
Hai CHENG ; Kai-Lin XU ; Hai-Ying SUN ; Qun-Xian LU ; Xu-Peng HE ; Xiu-Ying PAN
Chinese Journal of Hematology 2009;30(3):166-170
OBJECTIVETo construct a recombinant lentiviral vector (pXZ208-BDDhFVIII) mediating B-domain-deleted human coagulation factor VIII (BDDhFVIII) gene and investigate its expression in HLF, Chang-Liver and MSC cells.
METHODSBDDhFVIII gene fragment was separated by endonuclease digestion and was cloned into the multiple cloning sites of pXZ208 to construct a recombinant lentiviral vector pXZ208-BDDhFVIII. Viral particles were prepared by means of three-plasmid cotransfection of 293T package cells by calcium phosphate precipitation. After infection, the coagulant activity of human FVIII in the culture medium of 293T, HLF, Chang-Liver and MSC cells was assayed by one-stage method. The gene transduction efficiency was assayed by flow cytometry (FCM). Furthermore, PCR was performed to test the integration of BDDhFVIII.
RESULTSThe infection rates of HLF, Chang-Liver and MSC were (74.52 +/- 7.57)%, (27.24 +/- 6.53)% and (42.34 +/- 5.84)% respectively. The activities of FVIII in supernatants of HLF, Chang-Liver and MSC were (54.1 +/- 5.6)%, (22.5 +/- 2.9)% and (12.5 +/- 2.7)% respectively. BDDhFVIII gene integration was detected in all the infected cells.
CONCLUSIONThe recombinant lentiviral vector pXZ208-BDDhFVIII was successfully constructed and efficiently integrated into target cells to express human FVIII activity in vitro.
Cell Line ; Factor VIII ; biosynthesis ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Transfection
8.Association of rs4638289 and rs7131332 polymorphisms of the serum amyloid A1 gene with Kawasaki disease.
Ying CHEN ; Cheng WANG ; Qing JI ; Juan ZHANG ; Cheng TAN ; Shuang-Shuang WANG ; Xiu-Ying YI
Chinese Journal of Contemporary Pediatrics 2020;22(6):614-619
OBJECTIVE:
To study the association of the polymorphisms of the serum amyloid A1 (SAA1) gene at rs4638289 and rs7131332 loci with Kawasaki disease (KD) and its complication coronary artery lesion (CAL) in children.
METHODS:
A total of 105 Han children with KD who were hospitalized and treated from 2013 to 2017 were enrolled as the KD group. A total of 100 Han children who underwent physical examination were enrolled as the control group. According to the presence or absence of CAL, the KD group was further divided into a CAL group with 23 children and a non-CAL (NCAL) group with 82 children. Polymerase chain reaction-restriction fragment length polymorphism was used to investigate the polymorphisms of the SAA1 gene at rs4638289 and rs7131332 loci.
RESUKTS:
For the locus rs4638289 of the SAA1 gene, there were no significant differences between the KD and control groups in the genotype frequencies of AA, AT, and TT and the allele frequencies of A and T (P>0.05). But there were significant differences between the CAL and NCAL groups in the genotype frequencies of AA, AT, and TT (P=0.016), while there were no significant differences in the allele frequencies of A and T (P>0.05). AT genotype was a protective factor against CAL (OR=0.276, 95%CI: 0.099-0.772, P=0.011). For the locus rs7131332 of the SAA1 gene, there were no significant differences between the KD and control groups in the genotype frequencies of AA, AG, and GG and the allele frequencies of A and G (P>0.05). There were also no significant differences between the CAL and NCAL groups in the genotype frequencies of AA, AG, and GG and the allele frequencies of A and G (P>0.05).
CONCLUSIONS
Polymorphisms of the SAA1 gene at loci rs4638289 and rs7131332 are not associated with the onset of KD, while the polymorphism at the locus rs4638289 is associated with CAL in KD patients. KD patients with genotype AT may have a reduced risk of CAL.
Case-Control Studies
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Child
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Gene Frequency
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Genetic Predisposition to Disease
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Genotype
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Humans
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Mucocutaneous Lymph Node Syndrome
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genetics
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Polymorphism, Restriction Fragment Length
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Polymorphism, Single Nucleotide
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Serum Amyloid A Protein
;
genetics
9.The effects of interleukin-11 on high-dose methotrexate (HDMTX) induced mucositis in Wistar rats.
Yue-qin HAN ; Li-jun CHEN ; Xiao-jing SUN ; Guo-fa ZHAO ; Xiu-ying CHENG
Chinese Journal of Hematology 2004;25(12):740-744
OBJECTIVETo explore the therapeutic effect of interleukin-11 (IL-11) on high-dose methotrexate (HDMTX) induced mucositis in Wistar's rats, the proliferative effect on CEM leukemia cell line and the antitumor effect on HDMTX.
METHODSNinety-five 5-week old, 120 - 150 grams weight Wistar rats were randomly divided into five groups. Group A is normal control (n = 15), group B MTX control (n = 20), group C IL-11 pretreatment group before MTX injection (n = 20), group D (n = 20) the high dose IL-11 group (475 microg.kg(-1).d(-1)) after MTX injection, group E (n = 20) the low dose IL-11 group (150 microg.kg(-1).d(-1)) after MTX injection. All rats in group B approximately E were given 1 ml MTX intraperitoneally (100 mg/kg). Rats were killed at day 1, 3, 5, 7 after MTX injection. The mortality rates, changes of small intestine tissue morphology and ultra structure were observed. The proliferation of small intestine crypt cell was assayed by proliferating cell nuclear antigen (PCNA) immunohistochemical staining. MTT method was used to detect the proliferation of CEM cell line.
RESULTIL-11 treatment resulted in a significant increase of survival of HDMTX treated rats, increased of small intestinal villus length and villus/crypt ratio. IL-11 administration was associated with enhancement of small intestine mucosa recovery after HDMTX therapy. Group C showed a greater effect than group B (P < 0.01). IL-11 had no effect on CEM cell proliferation.
CONCLUSIONIL-11 has a significant mitigating effect on high-dose MTX induced intestinal mucositis in rat, and significantly increase the survival of the rats. IL-11 could be safely used in the HDMTX treatment of childhood acute lymphocyte leukemia.
Animals ; Antimetabolites, Antineoplastic ; toxicity ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Female ; Humans ; Immunohistochemistry ; Interleukin-11 ; pharmacology ; therapeutic use ; Intestinal Mucosa ; drug effects ; pathology ; ultrastructure ; Intestine, Small ; drug effects ; metabolism ; pathology ; Male ; Methotrexate ; toxicity ; Microscopy, Electron ; Mucositis ; chemically induced ; mortality ; prevention & control ; Proliferating Cell Nuclear Antigen ; analysis ; Random Allocation ; Rats ; Rats, Wistar ; Survival Rate
10.Study of the mechanical properties of Co-Cr ceramic alloy after recasts.
Song YANG ; Hui CHENG ; Xiu-rong LI ; Wei-qing WU ; Min ZHENG ; Ying-hui WANG
West China Journal of Stomatology 2011;29(3):249-252
OBJECTIVETo study the mechanical properties of Co-Cr ceramic alloy after recasts.
METHODSCo-Cr ceramic alloy cast samples were prepared and recast for 3 times without adding any new Co-Cr ceramic alloy. The tensile strength, 0.2% yield strength, percentage of elongation, flexural strength, flexural modulus and Vickers hardness of each specimen were measured.
RESULTSBeing cast for different times, the Co-Cr ceramic alloy showed no significant differences on their tensile strength, 0.2% yield strength, percentage of elongation, flexural strength, flexural modulus and Vickers hardness.
CONCLUSIONCo-Cr ceramic alloy can be recast for 3 times at least, without decrease of the mechanical properties.
Alloys ; Ceramics ; Chromium Alloys ; Dental Stress Analysis ; Hardness ; Materials Testing ; Metal Ceramic Alloys ; Tensile Strength