3.Effect of NGF on the sperm motility of human in vitro
Kai LIN ; Cuige SHI ; Yongqi ZHAO ; Shuhong LIU ; Ming FAN
International Journal of Laboratory Medicine 2012;33(20):2433-2437
Objective Motility is an important physiological characteristic of a mature sperm.Nerve growth factor(NGF) is a protein essential for the development,maintenance and survival of the peripheral and central nervous systems.NGF and its receptors TrkA and p75 are widely expressed in the testis,accessory reproductive organ,and the epididymal sperms.In the present study,we investigated the role of NGF on human sperm motility.Methods Use 0.1,1 and 10 μmol/L concentrations of NGF,on sperm motility study to investigate the optimal concentration.Use CASA to detect Sperm motility changes every 10 minutes in an hour after 10 μmol/L NGF was added to the semen.Results The parameters of sperm motility increased after NGF incubation had significant difference, in particular,VAP,VSL,VCL,BCF and LIN mean were significantly increased more than 32%.MAD,STR,ALH and WOB mean had no notable difference.Furthermore,NGF promotes the sperm motility in a time- and dose- dependent manner.In addition,the enhancement of NGF on sperm motility was more stronger than those of sperm culture medium.Conclusion Our findings suggest that NGF plays a promoted role in human sperm motility.
4.The role of membrane complement regulatory protein in transplantation of bone marrow mesenchymal stem cell
Kai XIAO ; Lin YANG ; Jingjing ZHAO ; Ruokun HUANG ; Ming XIE ; Kun LI
Chinese Journal of Orthopaedics 2016;36(5):277-282
Objective To explore the effect of complement activation on bone marrow mesenchymal stem cells (BMSCs)and evaluate the effect after transfection of complement regulatory proteins.Methods Bone marrow aspirate was harvested from 10 cases of patients suffered from fractures.Mesenchymal stem ceils were isolated,indentified cultured and then experimented in vitro.The complement cytotoxicity on the mesenchymal stem cells in autologous serum was measured by Europium cytotoxicity assay.The samples were divided into BMSCs group,BMSCs+ autologous human serum (AHS) group and BMSCs+ inactivated autologous human serum (iAHS) group.The complement membrane attack complex (MAC) deposited on the membranes was detected by flow cytometry.Finally,the cytotoxicity on BMSCs was measured after transfected with membrane complement regulatory proteins (mCRPs).All samples were divided into BMSCs with mCRPs untransfected group and BMSCs with mCRPs transfected group.Results More than 95% of cells derived from bone marrow were identified to be mesenchymal stem cells through detection of cell surface markers by flow cytometry.The cytotoxicity of untreated cells was 0.41%± 1.48%.BMSCs harvested from the 10 patients all had cytotoxicity after incubated with autologous serum,and the cytotoxicity was 32.59%±2.73%,while cytotoxicity after incubated with complement inactivated autologous serum was 2.59%±3.08%,which was similar to control group.Complement attack complex (MAC) could be detected on the BMSCs incubated with autologous serum,which implied the complement activation.After transfection of mCRPs,the cytotoxicity of autologous serum on transfected cells was decreased.The cytotoxicity of untransfected cells (41.70%±4.47%) had significant difference compared to the cells transfected with CD55 (21.87%±2.19%),the cells transfected with CD59 (18.67%± 1.42%),and the cells transfected with CD46+CD55+CD59 (28.43%±2.14%).CD55,CD59 and CD46+CD55 +CD59 transfected groups could impair effectively the cytotoxicity from complement.However,the cytotoxicity impairment was less effective in CD46 transfected cells (39.30%±3.96%),which had no significant difference compared to untransfected cells.Conclusion Membrane complement regulatory proteins could effectively protect bone marrow mesenchymal stem cells from attacks by complement.
5.Effect of tetrandrine eye drops on expression of apoptotic protein and transforming growth factor-β2 in human Tenon capsule fibroblast in vitro
Dai, LI ; Ming, SUN ; Qing-chun, LI ; Shao-chun, LIN ; Kai-li, WU
Chinese Journal of Experimental Ophthalmology 2012;30(3):218-222
BackgroundOur previous study determined that tetrandrine (Tet) has an inhibitory effect on the proliferation of human Tenon capsule fibroblasts ( TCFs ) in vitro,but its mechanism is poorly understood.ObjectiveThis study was to investigate the effect mechanism of Tet on human TCFs.MethodsHuman TCFs were isolated and cultured from scleral tissue of donor using explant technique.The cells were identified by vimentin antibody staining and morphology.The third generation of cells were seeded in the culture plate at the density of 1 × 105 cells/ml.Twenty-four hours after inoculation,the Tet of 1 × 10-5 mol/L was added in the well of culture plate,and the cells cultured only in 1640 medium served as the control group.The apoptosis of the cells was assessed by TUNEL,and the expressions of bax,bel-2,transforming growth factor-β2 (TGF-β2 ) in TCFs were detected using immunochemistry.Results The cultured cells showed the features of the fibroblasts in shape with the positive response for vimentin.A number of TUNEL positive cells were seen in Tet group and no TUNEL positive response was found in control group.The expression levels (A value) of bax,bcl-2 and TGF-~ protein in TCFs were 0.577 ± 0.009,0.430±0.012 and 0.341 ±0.017 in Tet group,and those in control group were 0.320±0.015,0.819±0.021 and 0.624±0.014 respectively,showing statistically significant differences between two groups( t =33.277,-35.356,-28.093,P<0.01 ).Conclusions Tet suppresses the proliferation of human TCFs through up-regulating the expression of bax and down-regulating the expressions of bcl-2 and TGF-β2 in vitro.
6.The correlation between ankle-brachial index and ultrasonography in lower extremities vascular in type 2 dia- betic patients.
Qing-Kai WANG ; Xue-Yan WENG ; Di-Ming ZHENG ; Ling LIN ; Al ET ;
Chinese Journal of Practical Internal Medicine 2006;0(S2):-
Objective To investigate the correlation between ankle-brachial index and ultrasonography in lower ex- tremities vascular in type 2 diabetic patients.Methods 218 type 2 diabetic patients were enrolled,and ankle-brachial index(ABI)measurements and uhrasonography in lower extremities vascular were performed in all patients.Lower extremi- ties vascular disease(LEVD)were divided into six species according to the uhrasonography results and scored in accord- ante with its serious degree,then investigate the correlation between ABI and uhrasonography results score.All patients were divided into deferent groups according to uhrasonography results and ABI range,and then compared their deference. Results The lower the ABI,the more serious of the LEVD showed by ultrasonography.ABI and ultrasonic inspection score was significantly negatively correlated(r=-0.65,P
8. ADAMTS9-AS1-SEMA3G affects the infiltration of immune cells in pancreatic cancer
Jia-kang MA ; Kai-kai REN ; Xiao-yan LIN ; Ming-yu HOU ; Bo ZHOU ; Jun MA
Journal of Medical Postgraduates 2020;33(2):169-173
ObjectiveThe mechanism that affects the infiltration of immune cells in pancreatic cancer has not yet been clarified. This study aims to investigate the lncRNA mRNA regulatory pathways that affect immune infiltration in pancreatic cancer.MethodsTCGA and GEO gene expression data were used to screen common differential lncRNAs. We perform survival analysis, target gene prediction, GO, KEGG enrichment analysis, immune infiltration analysis, gene set enrichment analysis (GSEA) on the selected differential lncRNAs to identify the relevant pathways of immune infiltration.ResultsThe pancreatic cancer patients with high expression of ADAMTS9 AS1 have a higher survival rate when compared to patients with low expression (P=0.010). The combined analysis of TCGA and GSE86436 revealed the difference and survival-related ADAMTS9 AS1. The functional prediction of ADAMTS9 AS1 was related to immunity. Using the TIMER database, the lncRNA affected the infiltration of immune cells in pancreatic cancer tissues. The clinical analysis was demonstrated that the ADAMTS9 AS1 was related to pathological grade. The target gene SEMA3G was screened by co-expression analysis using the IMMPORT database and TIMER database. Lastly, GSEA analysis of ADAMTS9-AS1 showed that the lncRNA was also related to tumor metabolism.ConclusionThese results indicate that ADAMTS9-AS1-SEMA3G is associated with the prognosis and immune invasion level of pancreatic cancer, which can provide a theoretical basis for subsequent genetic verification experiments and immune research.
10.Identification of chemical signals and haustorium induced of Thesium chinense.
Bo LIU ; Xiao-Ming ZHANG ; Qiao-Sheng GUO ; Chang-Lin WANG ; Lu CHEN ; Ling-Shan SONG ; Kai-Long AN
China Journal of Chinese Materia Medica 2014;39(23):4544-4547
To separate and identify chemical signals which induce Thesium chinense haustorium formation, the components of T. chinense roots secretion collected with XAD-4 resin were detected by GC-MS. The effect of DMBQ as exogenous signals to induce haustorium formation in T. chinense was studied. Fifty-three compounds of 9 types had been detected, including hydrocarbons, esters, organic acids, ketones, alcohols, nitrogen containing compounds, phenolic acids, aldehyde and quinine. It is worth noting that the 2, 5-di-tert-butyl-1,4-benzoquinone has the core structure of 1,4-benzoquinone, which may play an important role in the parasitic relationship of Prunella vulgaris and T. chinense: DMBQ worked effectively on inducing haustoria, but induction effects vary widely in different concentrations. DMBQ with the concentration of 1 μmol x L(-1) showed the best effect of the inducing ability with a ratio of 110.52 when treated to induce haustoria.
Gas Chromatography-Mass Spectrometry
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Host-Parasite Interactions
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Magnoliopsida
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chemistry
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physiology
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Plant Roots
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chemistry
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physiology
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Prunella
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chemistry
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physiology