1.Effects of diabetes on the expression and function of bradykinin 1 and 2 receptors after cerebral ischemia/reperfusion in rats
Hongfei SANG ; Zhongming QIU ; Ling LIU ; Lili XU ; Jun ZHANG ; Hao ZHANG ; Yi XIE ; Renliang ZHANG
Journal of Medical Postgraduates 2015;(5):458-464
Objective There is little research focusing on the expression and function of bradykinin 1 receptor ( B1R ) and bradykinin 2 receptor ( B2R) after cerebral ischemia/reperfusion on the basis of diabetes .The aim of this study was to compare the ex-pression difference and function change of B 1R and B2R in non-dia-betic and diabetic rats . Methods The cerebral ischemia/reperfu-sion model was established on 41 non-diabetic and type 2 diabetic rats, the weight and the biochemical index were measured on these two types of rats .8 non-diabetic rats and 8 diabetic rats were respec-
tively assigned to two groups according to random number tables:control group and I/R 24 h group, 4 in each group.Real-time PCR was performed to observe the expressions of two receptors at 24 h after reperfusion .Then, 33 non-diabetic rats and 33 diabetic rats were randomly divided into 4 groups respectively, including sham group (n=6), saline group (n=9), B1R antagonist group (n=9) and B2R antagonist group (n=9).At 24 hours after cerebral I/R, neurological deficiency was evaluated by neurological severity scores ( NSS);infarct volume was observed by TTC staining;cell apoptosis was determined by TUNEL staining;neuron degeneration was de-tected by Fluoro-Jade C staining. Results Glucoses of diabetics at 3, 7, 14 d after model establishment [(23.45 ±5.01), (23.71 ±4.87), (22.72 ±4.11) mmol/L] were obviously elevated compared with non-diabetics [(5.77 ±0.75), (6.05 ±0.69), (7.15 ±1.09) mmol/L];blood cholesterin [(4.59 ±3.43) mmol/L] and insulin [(67.26 ±12.02) pmol/L] at 14 d after model establishment were evidently incresaed in comparison to those in non-diabetics [(1.58 ±0.37) mmol/L, (25.34 ±4.88) pmol/L] (P<0.05), while no significant difference was found in the blood triglyceride of diabetics between them (P>0.05).Compared with non-diabetics, diabetics suffered from more apparent up-regulation of B1R mRNA (P<0.01) but relatively less B2R mRNA (P<0.05) at 24 h after I/R.NSS score, infarction volume, damaged and apoptotic cells in B2R antagonis-treated non-diabetic rats at 24 h after I/R conspicuously decreased compared with saline-treated non-daibetic rats.Those indicators in B1R antagonis-treated diabeics were strikingly lessened compared with saline-treated daibetics . Conclusion I/R induced distinct up-regulation of B2R mRNA in non-diabetics and inhibiton of B 2R effectively ameliorated the infarct volume and cell injury after I/R in non-diabetics; I/R induced more notable up-regulation of B1R mRNA in diabetics and B1R antagonist exerted neuroprotective effects instead of B 2R antagonist af-ter I/R in diabetics.
2.Assessment of left ventricular volume and function in patients with left ventricular non-compaction by contrast-enhanced three-dimensional echocardiography
Linli QIU ; Mingxing XIE ; Xinfang WANG ; Qing LYU ; Ling LI ; Yali YANG ; Li YUAN ; Zhenxing SUN
Chinese Journal of Ultrasonography 2014;(11):921-924
Objective To evaluate the value of echo‐contrast RT‐3DE for assessment of left ventricular volume and function in patients with left ventricular non‐compaction(LVNC) .Methods Twenty‐one patients of LVNC were involved and underwent non‐enhanced and contrast‐enhanced RT‐3DE to evaluate left ventricular end‐diastolic volume (LVEDV) ,left ventricular end‐systolic volume (LVESV) ,left ventricular ejection fraction (LVEF) .The endocardial border definition of LV was graded for each of the 16 LV segments as follows :0 = border invisible ,1 = border visualized only partially ,and 2 = complete visualization of the border .Three image‐quality groups (good ,fair ,and uninterpretable) were identified . Results ①Duringcontrast‐enhancedRT‐3DE,ascomparedwithnon‐enhancedRT‐3DE,thenumberof segments with complete visualization of the endocardial border increased significantly (55% vs 82% ,P <0.01) ,and the number of patients with a good‐quality echocardiogram increased significantly (33% vs 81% , P <0.01) .②Contrast‐enhanced RT‐3DE provided significantly larger values of LVEDV ( P < 0 0.1) and LVESV ( P < 0 0.1) as compared with non‐enhanced RT‐3DE ,the values of LVEF were not statistically different between the two techniques ( P =0.07) .③Intra‐and inter‐observer agreement for assessment of LV volumes and systolic function improved during contrast‐enhanced RT‐3DE ,as compared with non‐enhanced RT‐3DE .Conclusions Contrast‐enhanced RT‐3DE can increase the prevalence of good‐quality echocardiograms and significantly improve the reproducibility of LV volumes and function measurements .
3.Lead Exposure of Blood Source in Blood Change Treatment on Newborns
ling, XIE ; qiu-guo, CHENG ; li-ya, MO ; cai-zhi, HUANG ; bin, HU ; xun-liang, JIANG
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To explore the danger of lead exposure in newborns who accepted the blood stored in blood bank for blood change treatment.Methods The lead level of blood was examined before and after blood change treatment for 37 neonates with hyperbilirubinemia who accepted 53 cases blood stored in blood bank during Jun.to Dec.2006.The level of blood lead was measured by graphite stove atom absorb spectrum method.Results The average lead level of 53 cases blood stored in blood bank was 101.02 ?g/L,which had attained the level of lead poisoning.There were 15 cases(28.5%) whose blood lead levels was very high(≥100 ?g/L),3 cases whose blood lead level ≥200 ?g/L.After blood change treatment,the percentage of the blood lead level ≥100 ?g/L rose from 2.9% to 19.0%.The average level of blood lead after blood change treatment was higher than before(P
4.Expression of Recombinant sPDGFR?-Fc in CHO and Its Anti-proliferation Analysis
Yan WAN ; Li-Ling LI ; Qiu-Ling XIE ; Shu-Jun GUO ; Li QIN ; Yong-Cang ZHANG ; Xiao-Jia CHEN ;
China Biotechnology 2006;0(07):-
Orjective:To obtain recombinant CHO-K1 with expressing sPDGFR? and to identify the biological activities of sPDGFR? secreted in non-serum medium.Methods:Recombinant human sPDGFR? expression vector pIRES-Neo3-sPDGFR?-Fc was constructed and then transfected into CHO-K1 cells by using LipofectamineTM 2000.After screened with G418 in 8 weeks,some monoclone cells were selected randomly to amplify in 96-well-plate to 24-well-plates,and then to identify positive cell clones by RT-PCR.Furthermore,the candidate cell clones were test by Real-Time PCR and Western blot assays.Finally,anti-proliferation activities of the expressed sPDGFR? were analyzed by MTT.Results:sPDGFR?-Fc was cloned into pIRES-Neo3 correctly.The sPDGFR?-Fc expression level in recombinant CHO-K1 cell clones were concordant in between Realtime PCR and Western blot assay.sPDGFR?-Fc obtained from cultured non-serum medium of positive CHO-K1 could significantly inhibit proliferation of vascular endothelial cell.Conclusion:Successed to select recombinant CHO-K1 cell lines with high expressed sPDGFR?-Fc.The sPDGFR?-Fc can inhibit the cell proliferation significantly and it means sPDGFR?-Fc might be a new anti-cancer drug in the future.
5.Preparation and biocompatibility evaluation of osteochondral composite scaffold.
Qiu-ling WANG ; De-ming XIE ; Yu-ming LV
Journal of Southern Medical University 2010;30(11):2477-2481
OBJECTIVETo prepare osteochondral composite scaffold and study its biocompatibility in vitro.
METHODSThe composite material of nano-HAP/collagen I was prepared, and the osteochondral scaffold was manufactured by combining nano-HAP, collagen I, and PLGA as the bone section and sodium hyaluronate and PLGA as the chondral section. The diameter, chemical composition and crystallinity of the nano-HAP/collagen I composite particles were assessed with TEM, FTIR and XRD, and the biocompatibility and cytotoxicity of the scaffold were evaluated using MTT assay by co-culturing bone marrow stem cells and the scaffold.
RESULTS AND CONCLUSIONThe osteochondral composite scaffold has good microstructure without obvious cytotoxicity, possesses good biocompatibility with bone marrow stem cells and is suitable as an osteochondral scaffold material.
Biocompatible Materials ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Chondrocytes ; cytology ; Durapatite ; Humans ; Materials Testing ; Mesenchymal Stromal Cells ; cytology ; Tissue Engineering ; methods ; Tissue Scaffolds
6.Development and application of communication materials for participatory health education of schistosomiasis in fishermen and boatmen of Poyang Lake region
Jing ZHANG ; Zhaojun LI ; Ling QIU ; Dong LI ; Junjiang CHEN ; Hua XIE ; Yuemin LIU ; Fei HU ; Min YUAN ; Dandan LIN
Chinese Journal of Schistosomiasis Control 2016;(1):58-61
Objective To design and develop new types of health education materials which are suitable for fishermen and boatmen in endemic areas of marshland and lake regions,and to observe their application effects. Methods A total of 292 adult fishmen and boatmen who lived in Houshan Village,Yugan County,the schistosomiasis endemic area of Poyang Lake Re?gion,were selected randomly and investigated by questionnaires to understand the status of their knowledge,attitudes,practic?es on schistosomiasis control as well as the channels for getting information on schistosomiasis control and the materials that they were willing to accept. Then the information and materials suitable for the target population were developed together by the re?searchers and the volunteers of the villagers through focus group discussions,personal interviews and the Delphi method. Re?sults A series of participatory health education materials of schistosomiasis control targeted to the fishmen and boatmen were developed,including 2 live posters,2 picture puzzles,2 short opusculums and one song about schistosomiasis control. The field application showed that 98.97%,84.38%,78.35% and 80.93% of the participants considered those materials had scientificity, intelligibility,interestingness and practicability,respectively. Conclusion The participatory health education materials of schistosomiasis control is suitable for fishmen and boatmen,which can be used for reference by other endemic areas in marsh?land and lake regions.
7.Biological Characteristics of Mesenchymal Stem Cells Cultured in Conditions for Spermatogonia in Vitro
qiu-ling, TANG ; hong-yan, HE ; guang-yu, LIN ; li-min, LIN ; qing-dong, XIE ; tian-hua, HUANG ; lian, MA
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To investigate the possibility of inducing mesenchymal stem cells(MSCs)from human umbilical cord Wharton's Jelly to differentiate into spermatogonia.Methods To isolate,culture and purify MSCs with adherent method,the growth and proliferation of human umbilical cord-derived MSCs were observed,and their immunophenotypes were determined by flow cytometry;MSCs of the third generation were divided into 2 groups to be induced and cultured,MSCs of the control group were cultured in basal medium,while those of the experimental group with conditional medium.The morphologic and ultrastructure changes of control group and experimental group cells were compared with phase contrast microscopy,electron microscopy(EM)and transmission electron microscope(TEM)respectively ;the spermatogonial cells differentiated were then evaluated by immunohistochemistry stained for CD117and CD49f ;the method of Western-blot was used to test if the cells induced could express CD49f.Results A population of MSCs were isolated from human umbilical Wharton's Jelly;they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling;After induction,the shape of MSCs changed greatly from the fibroblast to the round,even familiar to the tadpole;expressed the known molecular markers of spermatogonial cells,such as CD49f,CD117.Conclusion The induced MSCs not only undergo spfermatogonial-cell like morphologic changes,ultramicrostructure mature with increasing cell organs,but also express the spermatogonial cell markers,which show that human umbilical cord derived MSCs are capable of differentiating into spermatogonial cell.
8.Study on Transplantation of Human Umbilical Cord Mesenchymal Stem Cells into Infertile Mouse Testis
qiu-ling, TANG ; guang-yu, LIN ; li-min, LIN ; tian-hua, HUANG ; qing-dong, XIE ; lian, MA
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To observe clone ability of human umbilical cord mesenchymal stem cells (MSCs) into infertile mouse seminife-rous tubules and the effects of MSCs on reproductive function.Methods Busulfan was used to destroy endogenous spermatogenesis of the recipient mice.To isolate,culture and purify MSCs with adherent method before marked with Brdu and Hoechst 33258 respectively,and then transplanted into the seminiferous tubules by microinjection.The survival of MSCs in recipient testes were evaluated by immunohistochemistry stained for Brdu and Fluorescent microscopy for Hoechst 33258 observation at different times.The diameter of seminiferous tubules was detected with HMIAS-2000 high-definition colored analyzing system for medical pictures.SPSS 13.0 software was used to analyze the data.Results The dosage of Busulfan resulted in 15% death in the mice,the testis of survived mice showed only basilar membrane in seminiferous tubules after 4 weeks.A lot of purified MSCs were obtained at the third generation and transplantation them into mouse seminiferous tubules survive for at least 4 months and appear to migration.The average diameter in experimental groups were higher than those in controls not only on 26 days but also on 120 days(P
9.Strong Expression of Recombinant Human Morphogenetic Protein-4 in Escherichia coli and its Bioassay in vivo
Si-Hong GAO ; Ju WANG ; Que-Wei DONG ; Kan LIU ; Xue-Ting LIU ; An HONG ; Qiu-Ling XIE ; Fen-Yong SUN ;
China Biotechnology 2006;0(03):-
Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.
10.Effect of L-methionine on trace elements in lead-intoxicated mice.
Ling-lin XIE ; Chang-hong ZHU ; Wei-qun TIAN ; Qiu-hua GAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(2):108-110
OBJECTIVETo investigate the effect of L-methionine (L-Met) on the content of Zn, Cu, Mn, Fe in liver, brain, spleen and kidney of lead intoxicated mice.
METHODSDistilled water was given to 10 mice (normal control group) and lead acetate solution of 400 micro g/ml Pb(2+) to 20 mice to serve as drinking water for 10 days. The lead administration was then withdrawn and lead exposed mice were randomly divided into two groups: the lead control group took distilled water as drinking water for 4 weeks to serve as positive control, the other one took L-Met solution (0.5 mg/ml) as drinking water for 4 weeks (Pb + L-Met group) to serve as the treatment group. All the animals were sacrificed on the 1st day after 4 weeks, and the contents of Zn, Cu, Fe, Mn, Pb in liver, brain, spleen and kidney were measured by Inductively Coupled Plasma (ICP) Emission Spectrometry.
RESULTSLead contents in liver, brain, spleen and kidney of Pb control group [(1.490 +/- 1.654) micro g/g, (3.470 +/- 2.757) micro g/g, (4.975 +/- 2.993) micro g/g, (0.066 +/- 0.001) micro g/g respectively], were higher than those in normal control group [(0.015 +/- 0.001) micro g/g, (0.009 +/- 0.007) micro g/g, (0.027 +/- 0.002) micro g/g, (0.006 +/- 0.015) micro g/g, P < 0.05] while Zn contents in liver, brain, spleen and Fe and Mn content in liver, brain, spleen and kidney in Pb control group were lower than those in normal control group (P < 0.05). Pb contents of brain, spleen and Cu content of kidney in Pb + L-Met group were higher than those in normal control group (P < 0.05). Zn contents of liver, brain, spleen, Fe contents of liver, brain, spleen, kidney, and Mn contents of brain, spleen in Pb + L-Met group were lower than those in normal control group (P < 0.05). Fe contents of liver, brain, Zn content of spleen, Cu content of kidney and Mn contents of liver, brain, spleen in the Pb + L-Met group were higher than those in the Pb control group (P < 0.05). The lead levels of four organs in the Pb + L-Met group were lower than those in the Pb control group (P < 0.05).
CONCLUSIONLead could be eliminated by L-Met, which may affect the distribution and metabolism of trace elements in mice.
Animals ; Brain ; metabolism ; Female ; Kidney ; metabolism ; Lead Poisoning ; metabolism ; Liver ; metabolism ; Male ; Methionine ; pharmacology ; Mice ; Spleen ; metabolism ; Trace Elements ; metabolism