1.Effect of hypoxia on expression of angiopoietin-2 in human retinal pigment epithelium
Chinese Journal of Experimental Ophthalmology 2012;30(9):825-829
Background Choroidal neovascularization(CNV)has been descrihed as a main reason of visual loss in a lot of ocular diseases.Researches showed that local hypoxia and retinal pigment epithelial(RPE) cells play an important role in the formation of CNV.A closely relationship of angiopoietin-2 (Ang-2) and angiogenesis has been proved.However,whether the expression of Aog-2 in hypoxic cultured human RPE cells is associated with the pathogenesis of CNV is still below understood.Objective This study was to investigate the effects of hypoxia on expression of Ang-2 in cultured human RPE cells in vitro,and discuss the possible effects of Ang-2 in the formation of CNV.Methods Human RPE cells were cultured and passaged,and 4-7 generation of cells were used in the experiment.The cells were incubated in cultural plate at the density of 5×107 cells/L.The culture medium containing 200 μmol/L CoCl2 was used to establish the hypoxia model of human RPE cells culturecd in vitro for 0.5,1,2,4,6,12and 24 hours,and the RPE cells cultured under normoxia were as controls.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of Ang-2 mRNA in cultured human RPE cells,and enzymelinked immunosorbnent assay(ELISA) was used to assay the content of Ang-2 protein in supernatant of cultured human RPE cells.Results The survival rate of human RPE cells was 90% after resoscitation.The fourth generation of cells showed the fusiform with the less pigment in them.The Ang-2 mRNA/β-actin mRNA value in human RPE cells was significantly different among various groups(F=1086.30,P=0.00),The Ang-2 mRN A/β-actin mRNA value in hypoxia cultured for 0.5 hours group began to increase and peaked in hypoxia culture for 4-6 hours group,with the significant differences in comparison with normoxia control group(P<0.05).The Ang-2 mRNA/β-actin mRNA value decreased to the baseline level at hypoxia for 24 hours.The ELISA analysis showed that the concentration of Ang-2 protein in supernatant of RPE cells showed significant difference among groups(F=1034.00,P=0.00).The concentration of Ang-2 protein increased at hypoxia culture for 0.5 hours and peaked at 6 hours,showing significant differences in comparison with the control group (P<0.05).Conclusions Hypoxia could significantly up-regulate the expression of Ang-2 in human RPE cells cultured in vitro.Ang-2 expresses highly in the early stage of hypoxia,implying that Ang-2 participates in the formation of CNV.
2.Autogenous peripheral stem cell transplantation combined with interleukin-2 for treatment of 16 patients with lymphoid malignancies
Yanhui XIE ; Qinfen CHEN ; Pei LI
China Oncology 2001;0(05):-
Purpose:To evaluate the effect of Auto-PBSCT combined with long-time low-dose interleukin-2 (after transplant) for increasing the rate of 5-year disease-free survival of lymphoid malignancies (ALL and NHL).Methods:U- sing chemotherapy combined with low dose G-CSF to mobilize peripheral blood stem cells,we carried out auto-PBSCT in lymphoid malignancies (7 cases CR_1 ALL、1 case CR_2 ALL、3 cases CR_1 NHL、5 cases refractory NHL).After transplant, 15 cases used low dose interleukin-2 at 5.10~5U/d for one year or longer.Results:7 cases CR_1 ALL are still alive,the aver- age disease-free survival is 73 months.Median disease-free survival is 51.5 months.2 of 3 cases CR_1 NHL are still alive. The 6 years disease-free survival in 10 cases CR_1 lymphoid malignancies(CR_1 NHL and CR_1 ALL) is 0.90?0.11,the aver- age disease-free survival is 66.8 months.In another PR (refractory) group of NHL and ALL (CR_2),3 years disease-free survival was 0.33?0.18,and average survival time was 20.66 months,but a chemotherapy-sensitive NHL is still alive (63 months),a case of lymphoblast cell NHL also survived for 46 months and died after relapse.Conclusions:Auto-PBSCT combined with long-turm low dose IL-2(after transplant) could effectively increase the rate of 5-year disease-free survival in CR_1 lymphoid malignancies(ALL and NHL).
3.Estimate a method of automatic extracting genomic DNA from large numbers of whole blood samples
Changrong CHEN ; Bin PEI ; Jinzhen XIE
Chinese Journal of Blood Transfusion 2002;0(05):-
Objective To estimate an automatic DNA extracting method, and evaluate the quantity and quality of these DNA samples.Methods Genomic DNA was extracted automatically from blood samples by MiniPrep 75-Ⅱ workstation. The yield and purity of DNA samples were detected by UV-spectrophotometer, and the integrality of these DNA samples were measured by agarose gel electrophoresis.Results The yield of genomic DNA extracted from 100?l whole blood were (7.33?2.58)?g. The average purity of these DNA samples was 1.647?0.135 (A260/A280) and the molecular weight of them was about 21 kb.Conclusion High quality and intact genomic DNA can be extracted rapidly from whole blood by using MiniPrep 75-Ⅱ workstation, and these DNA samples were suitable for downstream molecular biology experiments.
4.Ultrasonic diagnosis of pediatric ovarian torsion
Xiaoying XIE ; Guanghua PEI ; Shicheng WANG
Chinese Journal of Ultrasonography 2015;24(2):144-146
Objective To evaluate the value of ultrasonography in the diagnosis of pediatric ovarian torsion (OT).Methods Ultrasonography was performed in 38 OT,and 52 non-OT children.Bilateral ovarian area was measured.The larger area was defined as S1,whereas,the smaller area was defined as S2.S1/S2 ratio was calculated.Follicles around ovaries,ovarian blood flow,and pelvic effusion were also observed during the ultrasonographic scanning.ROC curve was employed to evaluate the clinical value of ultrasonography in pediatric OT diagnosis.Results ①S1/S2 ratio,incidence of follicles around ovaries and of no ovarian blood flow were all higher in OT compared to non-OT children (P =0.000).While pelvic effusion showed no significant difference between the two groups (P =0.004).②Using S1/S2 ratio to diagnose OT,the area under the ROC curve was 0.925 (0.865-0.986),the best cutoff point was 1.6,sensitivity was 97% and specificity was 88%.Patients were divided into two groups based on the cutoff point of 1.6,the area under the ROC curve of S1/S2 was 0.924(0.847-0.969),of follicles around ovarian was 0.896(0.781-0.931),and of no ovarian blood flow was 0.911 (0.831-0.961),indicating S1/S2 held the highest diagnostic value.Conclusions Ultrasonography is an accurate method in the early diagnosis of pediatric ovarian torsion,with S1/S2 ratio holding the highest diagnostic value in this application.
5.Mitochondrial protein NDUFS3 and tumor
Pei WANG ; Xiaodong CHENG ; Xing XIE
Journal of International Oncology 2012;39(9):647-650
NDUFS3 is an essential subunit of mitochondrial NADH:ubiquinone oxidoreductase (complex Ⅰ ) and plays a critical role in the mitochondrial typeⅠ respiration chain.Mutations in this gene are shown to cause neurodegenerative disease such as Leigh syndrome (subacute necrotizing encephalopathy).In recent years,many evidences show that the expression of NDUFS3 proteins are lower in many cancerous cells compared to the corresponding normal cells.It comes to the conclusion that NDUFS3 may play a role in the tumorigenesis.
6.Study of the Biocompatibility Between Bone Morphogenetic Protein Microspheres and Bone Marrow Stem Cell
Yu QIN ; Guoxian PEI ; Deming XIE
Journal of Chinese Physician 2001;0(05):-
Objective To evaluate the biocompatibility between bone morphogenetic protein(BMP) composition and bone marrow stem cell (BMSc).Methods Microspheres were prepared with chitosan and sodium alginate, and BMP was enwrapped in the microspheres. The biocompatibility of the composition was examined using cell-culturing method. The BMSc was cultured in combination with microspheres. The extending speed of the cells, the proliferation and alkaline phosphatase activity were tested.Results There were no inhibition on cellular proliferation of BMSc when it was cultured in combination with microspheres in vitro, but ALP activity increased significantly.Conclusion BMP microspheres possessed satisfactory biocompatibility, and could increase the osteogenic capability of BMSc in vitro.
8.Mechanism of action of neuregulin protecting the myocardium against daunorubicin-caused damage in rats.
Sha-yi JIANG ; Pei-ran MA ; Xiao-tian XIE
Chinese Journal of Pediatrics 2006;44(7):541-543
Animals
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Antibiotics, Antineoplastic
;
administration & dosage
;
toxicity
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Apoptosis
;
drug effects
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Daunorubicin
;
administration & dosage
;
toxicity
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Disease Models, Animal
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Female
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In Situ Nick-End Labeling
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Male
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Myocardium
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cytology
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metabolism
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pathology
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ultrastructure
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Neuregulins
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metabolism
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pharmacology
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RNA, Messenger
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metabolism
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Rats
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Rats, Wistar
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Receptor, ErbB-2
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genetics
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metabolism
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Reverse Transcriptase Polymerase Chain Reaction
9.The effects of Ortho-K contact lens by day and overnight wear on wave-front aberrations
Dan WANG ; Pei-Ying XIE ; Zhi-Xin WANG ;
Ophthalmology in China 2006;0(05):-
Objective To investigate the change of wave-front aberrations after Ortho-K contact lens(CL)by day and overnight wear.Design Retrospective case series.Participant 180 eyes of 92 myopic patients wearing Ortho-K CL.Method Wave-front aberra- tions of 180 eyes of 92 myopic patients wearing Ortho-K CL were analyzed.Patients were divided into by day wear and overnight wear groups.Wave-front aberrations were measured with WFA 1000B subjective aberrometer before and 1 month after Ortho-K CL wear. Wave-front aberrations at initial,after one month of CL fitting and without CL were measured in same condition,and RMS values for overall wave-front aberrations and each order of the Zernike aberrations were analyzed with Matlab software.Main Outcome Measures RMS value of Zernike coefficient of each order.Result Compared with initial aberration in wear by day group,total RMS and each or- der aberrations(except 2nd order)with lens fitting significantly increased(P0.05).Compared with initial aberration in overnight wear group,overall wavefront aberrations and the RMS(except 2nd,3rd and 6th order Zernike aberrations)with lens fitting significantly increased(P0.05).Conclusion The high order aberrations with lens fitting and without lens fit- ting increased in each group.The aberration with lens fitting in wear by day group was higher than that in overnight wear group,while the aberration without lens fitting in overnight wear group was higher than that in wear by day group.(Ophthalmol CHN,2007,16: 351-354)
10.An experimental study of vascular endothelial growth factor and transforming gro wth factor a_1 expression in bone defect resulted from non-union
Xinlong MA ; Jun XIE ; Pei WANG ; Al ET
Chinese Journal of Orthopaedics 1996;0(09):-
Objective To observe the expression of vascular endothelia l growth factor (VEGF)and transforming growth factor a 1 (TGF-a 1 )in callus of non-union,bone defect for investigation of the molecular mechanisms of bone defect resulted from non-union,and further explore the co rrelative influences of VEGF and TGF-a 1 in fracture healing.Methods The bone defect resulted from non-union model and the fracture healing model were established in the radius of rabbit.The sp ecimens of callus and soft tissue were collected postoperatively at1,2,4, 6,8and12weeks.The expressions of VEGF and TGF-a 1 were in-vestigated by means of immunohistochemistry(SP method).Results There was no sign of subperiosteal healing except a large segment filled with hemorrhage and granulation tissue in bone defect area.There was only a little amount of granulation tissues transforming into chondrocytes.Some chondrocyt es stopped prolif-erating after reaching hyper-trophic chondrocyte stage but changed into fibrous connective tissues.Most of the granulation tissue finally changed to fibrous connective tissues in the fracture gap.In addition,there w ere a large number blood vessels invading the normal fracture healing site than that in non-union.In im muno-histochemistry observation,there were significa nt decreasing expressions of VEGF and TGF-a 1 in callus of different period of non-union comparing to union(P