ABSTRACT
Aim To explore the effect of Neu-P11,a novel melatonin agonist with similar function of melatonin,on IOP of acute high IOP animals and the related mechanism.Methods The experiment used the Trendelenburg position(head low feet high position of 80°)to establish acute high IOP model.Rats were placed in the Trendelenburg position and used Tonopen XL contact tonometer to measure IOP(every 5 minutes measured once IOP,and the maximum value in 20 minutes)in 8 :00～9 :00 am.And then,thirty Sprague-Dawley rats(8 week-old)were divided into five groups: normal IOP+normal saline,high IOP+normal saline,high IOP+10 mg·kg-1 Mel,high IOP+20 mg·kg-1 Neu-P11,high IOP+50 mg·kg-1 Neu-P11.Put in a flat to rest 2 h,animals were placed in Trendelenburg position again and then,IOP was measured every hour in the flat by 6 hours.After excessive sodium pentobarbital administration continuous for 1 week,the serum was collected and stored for subsequent detection at the end of the experiment.The level of MDA,SOD and GSH-Px enzyme activity of the rat serum was tested by kit accordingly.HE staining method was used to identify the SD rat retinal morphological changes.Results Trendelenburg position could induce IOP of model group rats,which was increased by 202.9％(P<0.01)and the content of MDA,reduced the activity of SOD and GSH-Px enzyme,retinal thickening was observed and its level was not clear.Neu-P11/Mel could significantly improve oxidative stress level and retinal edema in rats.Conclusion Neu-P11 could reduce IOP of the acute high IOP animals,which might be involved in the lower level of oxidative stress in the body.
ABSTRACT
Objective To explore the effects of the new melatonin nonselective agonists Neu-P11 on intraocular pressure (IOP) and glial fibrillary acid protein (GFAP) expression in the retina of acute high IOP rat.Methods Twenty-four male Sprague-Dawley rats were randomly divided into 4 groups (6 cases in each group):Normal IOP with local treatment (NIL) group,high IOP with local treatment (HIL) group,HILwith melatonin treatment (HIL-M) group,HIL with Neu-P11 treatment (HIL-N) group.10 μL normal saline was instilled in NIL group and HIL group,while 10 μL 100 μmol · L-1 Mel/Neu-P11 treated in HIL-M group and HIL-N group.After 2 hours of rest,rats were placed in the Trendelenburg position duration 45 minutes.And then,IOP was measured every hour for 6 hours,and repeated it for a week.The excessive sodium pentobarbital was injected to SD rats at the end of the experiment.The rat eyeballs were took out to perform HE and immunohistochemical staining to detect retina GFAP protein expression.Results After a week,IOP in HIL group was (41.26 ± 1.73) mmHg (1 kPa =7.5 mmHg),NIL group was (13.61 ± 0.55) mmHg,which mean the Trendelenburg could induce high IOP in SD rats.Compared with the NIL group,the retinal becoming thick,the level of organization was not clear and the expression of GFAP protein was quite high in HIL group.At the same time,the GFAP protein expression and IOP were significantly weakened in HIL-M group and HIL-N group compared with HIL group.Conclusion Neu-P1 1 can reduce IOP,inhibit the activation of gliocyte,and decrease the expression of GFAP to protect the retina.
ABSTRACT
Objective:To analyze the protective effects of adoptively transferring different patrilineal lymphocytes and their Exosomes ( Exo) on fetation of mice with pregnancy loss comparatively.Methods: The peripheral blood mononuclear cells ( PBMC) from healthy men and the splenocytes from BALB/c and DBA/2 male mice were induced in vitro ,and their Exo were isolated through sucrose gradient ultra-centrifugation combined with ultrafiltration.The mice of CBA/J (♀) mated with BALB/c (♂) were enrolled as control group of normal pregnancy ,and the CBA/J (♀) mated with DBA/2 (♂) as URSA of pregnancy loss experimental animal model.The mice in URSA group were randomly divided into each group with treatment through adoptively transferring , which were injected intravenously or subcutaneously with splenocytes or splenocytes -derived Exo from mated DBA/2,unmated DBA/2 or unrelated BALB/c,also PBMC-derived Exo from men,respectively.And then,the placenta volumes,rates of fetal absorption and pregnancy loss were calculated to observe the fetation of embryos.Results:Compared with the group of normal pregnancy ,the placenta volumes from URSA group decreased greatly ,and rates of fetal absorption and pregnancy loss elevated greatly ( all P0.05 ).After transferring the Exo derived from either male mice or healthy men,the level of decreased fetal absorption rates were more than that in cellular-therapy groups ( all P<0.05 ).After transferring the Exo derived from men ,the level of decreased pregnancy loss rates were more than that in cellular -therapy groups and mice splenocytes-derived Exo group ( all P<0.05 ).Conclusion:Adoptively transferring patrilineal T lymphocytes and their Exo can greatly improve the fetation.Exo should become a non-cellular bio-remedy,which is expected to replace traditional immunotherapy of adoptively transferring lymphocytes.
ABSTRACT
Objective To explore the effects and mechanisms of periostin overexpression on migration and invasion of nasopharyngeal carcinoma ( NPC) cell line.Methods The recombinant plasmids [ pCMV-neo ( +)-periostin ] and control plasmids [pCMV-neo (+)] were transfected into 6-10B cells using lipofectamine 2000TM reagent.The expression of periostin was detected with PCR and Western blotting .Transwell chamber invasion assay was employed to assay the migration and invasion of 6-10B cells before and after transfection .A gelatin zymogram was used to detect the activity of MMP-2 and MMP-9 in cultivated supernatant of 6-10B cells before and after transfection .The expression of integrin-αvβ5 was detected by immunohistochemistry ( IHC) in 6-10Bperiostin cells, 6-10Bvector cells and 6-10B cells as well as normal nasopharyngeal mucosa ( NNM) and NPC and at the same time periostin also was detected by immumohistochemistry in NNM and NPC, and densitometry analysis using image-pro plus 6.0 software, and the correlation between periostin and integrin-αvβ5 on NPC was assayed with statistics .Results Over expression of periostin promoted cell migration and invasion.The expression levels of integrin-αvβ5 in primary NPC and 6-10Bperiostin cells were significantly higher than those in NNM and 6-10Bvector, 6-10B cells.The expression in NPC of integrin-αvβ5 showed positively correlated with the expression of periostin (r=0.682, P<0.01).Conclusion Periostin plays an important role in regulation of cell migration and invasion probably by combining with integrin-αvβ5 to improve the activities of MMPs .
ABSTRACT
Objective To analyze the relationship between HBsAg level and HBV DNA level as well as illness severity in patients with HBV-related acute-on-chronic liver failure (ACLF).Methods One hundred and nineteen patients with chronic hepatitis B (CHB group) and 98 patients with ACLF(ACLF group) were enrolled.HBsAg and HBeAg were assayed with Roche electrochemical luminescence method.HBV DNA was quantified using a real-time polymerase chain reaction (PCR) assay.HBsAg and HBV DNA levels were compared between two groups and between HBeAg-positive and HBeAg-negative patients in ACLF group respectively,also the correlationbetween HBsAg and HBV DNA was studied.Results The proportions of HBeAg-negative patients were 68.4％(67/98) and 42.9％(51/119) in ACLF group and CHB group respectively,and there was significant difference between two groups (P ＜0.01).There was no significant difference in HBV DNA between two groups (P ＞ 0.05).HBV DNA in HBeAg-positive patients was higher than that in HBeAg-negative patients in two groups(P ＜ 0.05).There was no significant difference in HBsAg between HBeAg-positive patients and HBeAg-negative patients in ACLF group (P ＞ 0.05 ),but they were higher than that in HBeAg-positive patients in CHB group (P＜ 0.05).HBsAg was correlated to ALT,AST in HBeAg-positive patients (P ＜ 0.05).No significant correlation was found among HBsAg and HBV DNA as well as biochemical changes (P＞ 0.05).There was no significant difference in the ratio of different HBsAg levels among the patients of different HBV DNA in ACLF group (P＞ 0.05).Conclusion The level of HBsAg does not directly correlate with serum HBV DNA level,and has no directly correlation with the severity of the disease in patients with HBV-related ACLF.
ABSTRACT
OBJECTIVE@#To establish the protein expression map of nasopharyngeal carcinoma (NPC), and provide a basis for proteomic study of NPC.@*METHODS@#Laser capture microdissection (LCM) was used to isolate cancer cells from NPC tissues. Two-dimensional gel electrophoresis(2-DE) was used to separate the total proteins of LCM purified NPC cells. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) was performed to identify the proteins, and bioinformatics was used to construct the 2-DE database of NPC proteome.@*RESULTS@#A 2-DE reference map of NPC was established. On the 2-DE map, a total of (1 312+/-30) protein spots were detected, and 427 protein spots representing 241 non-redundant proteins were identified. The 2-DE database of NPC proteome was constructed. These data could be accessed at our website (http://www.xyproteomics.org).@*CONCLUSION@#A protein expression profile of LCM purified NPC tissues has been established for the first time, which provides useful information and source for proteomic study of NPC.
Subject(s)
Humans , Computational Biology , Gene Expression Profiling , Lasers , Microdissection , Methods , Nasopharyngeal Neoplasms , Genetics , Metabolism , Neoplasm Proteins , Genetics , Metabolism , Proteome , Metabolism , Proteomics , Methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
The mechanism of how stroma plays an important role in tumor carcinogenesis is now a hotspot. To delineate the features of stromal protein between nasopharyngeal carcinoma (NPC) and normal nasopharyngeal mucosa(NNM), laser capture microdissection (LCM) was performed to purify stromal cells from the NPC and NNM, respectively. The protein expressed profiles of the stroma of NPC and NNM were compared using fluorescent two-dimensional difference gel electrophoresis (2D-DIGE) and 34 differential protein spots between tumor stroma and normal stroma were chosen to be identified by mass spectrometry (MS). A total of 20 differential proteins were identified, and three differential proteins (CapG, L-plastin and S100A9) were selectively further validated by Western blotting and immunohistochemical analysis to confirm the results of 2D-DIGE. 2D-DIGE patterns of the stroma of NPC and NNM were established for the first time, the results suggested that differentially expressed proteins in the stroma of NPC and NNM may be useful for understanding the relationship between NPC cells and their surrounding microenvironment. Further studying of these proteins will be helpful to elucidate the mechanisms of NPC carcinogenesis and provide new thoughts on therapy of NPC through stroma.
ABSTRACT
To search for nasopharyngeal carcinoma (NPC) biomarkers,laser capture microdissection (LCM) was used to purify the target cells from NPC and normal nasopharyngeal epithelial tissues (NNET). Two-dimensional gel electrophoresis (2-DE) was performed to separate the total proteins of microdissected NPC and NNEC,PDQuest software was applied to analyze 2-DE images,and the differential protein spots between the two types of tissues were identified by both MALDI-TOF-MS and ESI-Q-TOF-MS. The expression of cytokeratin 8(CK8),one of the differential proteins,in the microdissected NPC and NNEC as well as 4 NPC cell lines with different differentiated degrees and/or metastatic potentials was detected by Western blot. Immunohistochemistry was also used to detect the expression of CK8 in paraffin-embedded tissues including 63 cases of primary NPC,28 cases of NNET and 20 cases of cervical lymphonode metastasis. In the present study,2-DE patterns of microdissected NPC and NNEC were established,and 29 differential proteins in the above two tissues were identified,of which 15 only expressed or up-regulated in NPC and 14 only expressed or up-regulated in NNET. The expression level of differential protein CK8 between the NPC and NNET was selectively confirmed,and was found to be related to the differentiation and/or metastasis of NPC cell lines. Significant down-regulation of CK8 was observed in NPC compared with NNET,and significant up-regulation of CK8 was also observed in lymphonode metastasis compared with primary NPC. The data suggest that CK8 may be related to the differentiation and lymphonode metastasis of NPC,and may serve as molecular biomarkers for metastasis and differentiation of NPC.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of nitric oxide(NO) on the DNA ploidy of germ cells and to evaluate the role of NO in modulating spermatogenesis by using SNP, a donor of NO and N-nitro-l-arginine-mythel-ester(L-NAME), an inhibitor of nitric oxide synthese(NOS) in rats physically in vivo.</p><p><b>METHODS</b>Forty adult male, Sprague-Dawley rats (60-70 days) were divided into four groups, and injected ultraperitoneally with one of the following agents (once a day, for 12 days): SNP, L-NAME and SNP + L-NAME with normal saline. Two hours after the last injection the rats were sacrificed. The sera were collected and stored at -70 degrees C for subsequent hormone assay. The concentration of serum testosterone was measured by radioimmunoassay. Serum NOx- (nitrite/nitrate) concentration was measured by Greiss method. DNA of spermatogenic cells was detected by flow cytometry(FCM), and the percentage of 1c, 2c and 4c germ cells calculated.</p><p><b>RESULTS</b>In the SNP treatment group, the serum concentration of NOx- was higher, testosterone concentration was lower and the number of 1c cells was smaller compared with the control group. However, in rats treated with L-NAME, the concentration of NOx- was significantly lower, testosterone concentration was higher and the number of 1c cells was larger compared with the control group(P < 0.01). No changes were observed in the SNP + L-NAME group.</p><p><b>CONCLUSION</b>Enhancing ectogenous NO will suppress spermatogenesis while inhibiting NO productive pathway will promote it.</p>
Subject(s)
Animals , Male , Rats , DNA , Flow Cytometry , NG-Nitroarginine Methyl Ester , Pharmacology , Nitric Oxide , Physiology , Nitroprusside , Pharmacology , Rats, Sprague-Dawley , SpermatogenesisABSTRACT
Objective To evaluate the effect of sodium nitroprusside(SNP)and N-nitro-l-arginine-mythel-ester (L-NAME) on apoptosis of spermatogenic cells in rats. Methods Fourty adult male Sprague-Dawley rats (60-70days) were divided into four groups.Each group was injected intraperitoneally with one of the following agents, once a day, for 12 days: 1. SNP; 2.L-NAME;3.SNL+L-NAME;4.Normal saline NS group.Two hours after the last time injection the rats were sacrificed.TUNEL staining and flow cytometry analysis were used to detect the apoptosis of spermatogenic cells. Results Sub-monoploid and apoptosis index (AI) in SNP group was significantly higher than that of NS group and sub-monoploid and apoptosis index (AI) in L-NAME group were significantly lower than that of NS group by FCM and TUNEL (P0.5) was found.Conclusion SNP can accelerate the apoptosis of spermatogenic cells and L-NAME can inhibite the apoptosis of spermatogenic cells,The effect of SNP and L-NAME on apoptosis of spermatogenic cells probably occurs through the action of nitric oxide.