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1.
Article in Chinese | WPRIM | ID: wpr-451820

ABSTRACT

AIM:To explore the effects of pGRIM-19-si-survivin co-expression plasmid carried by human at-tenuated Salmonella on prostate cancer subcutaneous xenograft growth in nude mice .METHODS: Prostate cancer xeno-graft model was established in nude mice .Co-expression plasmids carried by attenuated Salmonella were introduced by in-traperitoneal injection .The xenograft volumes were monitored timely .Immunohistochemical staining , RT-PCR and TUNEL assay were applied to investigate the related mechanisms that pGRIM-19-si-survivin inhibited tumor growth in vivo.RE-SULTS:Compared with psi-survivin and pGRIM-19 carried by attenuated Salmonella ( control groups ) , the tumor volumes were reduced markedly in pGRIM-19-si-survivin plasmid group .The mean shrinkage rates were 2.36 and 3.02 times. pGRIM-19-si-survivin co-expression plasmid carried by attenuated Salmonella inhibited survivin expression but strengthened GRIM-19 expression obviously (P<0.05).The mRNA expression of apoptosis-related proteins such as Bcl-xL, Stat3, cy-clin D1 and c-Myc was inhibited , and the vascular endothelial growth factor ( VEGF) mRNA and Ki67 protein were also in-hibited, but the caspase-3 mRNA expression was up-regulated ( P<0.05 ) with significant cell apoptosis .CONCLU-SION:pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella inhibits the growth of prostate cancer subcutaneous xenografts in nude mice by promoting cell apoptosis and inhibiting prostatic cancer proliferation .

2.
Basic & Clinical Medicine ; (12): 139-143, 2010.
Article in Chinese | WPRIM | ID: wpr-440596

ABSTRACT

Objective To investigate the role of human membrane associated sialidase (Neu3) in multidrug resistance ( MDR) of K562 cells. Methods K562 cells and K562/ADM cells were treated with DNR alone or with combination of DNR and NeuAC2en. The cell survival rate was measured by MTT assay; the expression of MDR related factors and apoptosis related factors were determined by Western blot and RT-PCR; Neu3 activity was detected by TBA reaction. Results The survival rate of K562/ADM cells was higher than that of K562 cells; NeuAC2en showed synergistic effect with DNR(P <0. 01) ; Neu3 activity of K562 and K562/ADM cells decreased after DNR or NeuAC2en induction and the decrease was most significant in the combination treatment group (P <0. 01). Under the identical condition, the protein expression of P-gp and Neu3 and the mRNA expression of MDR1, Neu3, BCL-xl and BCL-2 increased comparing with K562 cells. The mRNA level of BAX in K562 and K562/ADM groupsdid not changed significantly. MDR1, Neu3,BCL-xl and BCL-2 decreased after DNR induction and down-regulated most obviously in the groups treated by DNR combined with NeuAC2en. Conclusion Neu3 may be related with multidrug resistance of K562/ADM cell through regulating apoptosis and the expression of MDR1.

3.
Chinese Journal of Pathophysiology ; (12): 2136-2140, 2009.
Article in Chinese | WPRIM | ID: wpr-405492

ABSTRACT

AIM: Aquaporin 3 ( AQP3 ) water channel expressed in the kidney plays a critical role in the urine concentrating mechanism. Mice with AQP3 deletion show a urinary concentrating defect. To better characterize this defect, we studied the influence and mechanism of an acute urea load in conscious AQP3 - null and wild - type mice. METHODS:Urine was collected and assayed every 2 h, from 2 h before (baseline) to 8 h after the urea load. Urine volume, urine osmolality and urea concentration were measured. RNA was isolated from the whole kidney and real - time PCR was carried out using a LightCycler. Total protein of UTs was assayed from kidney tissue by Western blotting. RESULTS: Mice of both genotypes excreted the urea loaded in ~ 4 h with the same time course. Despite their low baseline, the AQP3 - null mice raised their urine osmolality and urea concentration progressively after the urea load to the values almost equal to those in wild - type mice at 8 h. In contrast, urine non - urea solute concentration did not change. Urine volume fell in the last 4 h to about one - fourth of basal values. The urea load strongly upregulated urea transporter UT - A3 expression in both genotype mice. CONCLUSION: These observations show that lack of AQP3 does not interfere with the ability of the kidney to concentrate urea but impairs its ability to concentrate other solutes. This solute - selective response results from the capacity of AQP3 to transport not only water but also urea, suggesting a novel role for AQP3 in non - urea solute concentration in the urine.

4.
Article in Chinese | WPRIM | ID: wpr-596505

ABSTRACT

Objective To investigate the effects of polyi:c on angiogenesis in mouse prostate carcinoma tissue and their possible mechanisms.Methods Mouse prostate carcinoma models were randomly divided into two groups according to tumor weight:control group and polyi:c group.After treatment for 7 times,the mice were sacrificed and the tumor tissues were cut for weighing,calculating the tumor inhibitory rate and tumor index.Hematoxylin-eosin staining and immunohistochemical staining were performed to observate the morphological changes of prostate carcinoma tissues,distribution of vasa and expressions of vascular endothelial growth factor (VEGF) and endothelial nitric-oxide synthase (eNOS).Results In polyi:c group,the mean tumor inhibitory rate was 67.85% and the tumor index was(5.42?0.17)%;in control group,the tumor index was(14.45?1.06)%; there was significant differences between polyi:c group and control group( P

5.
Article in Chinese | WPRIM | ID: wpr-592688

ABSTRACT

Objective To investigate the role of the mass screening by comparing the pathological features of prostate cancer between mass screening patients and clinical patients.Methods 107 cases of prostate cancer(including 51 patients from clinical diagnosis and 56 patients from mass screening) and 7 cases of prostate intraepithelial neoplasia(PIN,from mass screening) were analyzed using the Gleason’s grade system.Results ① Gleason’s grade of prostate cancer in mass screening group was lower than that in clinical diagnosis group(?2 =48.22,P

6.
Article in Chinese | WPRIM | ID: wpr-591788

ABSTRACT

Objective To investigate the alteration of aquaporin 1(AQP1) expression in lung tissues in hemorrhage-lipopolysaccharide(LPS) two-hits rats and the effects of panaxadiols(PDS)and dexamthasone(Dex) on it.Methods The rat model of acute lung injury was built with hemorrhage-LPS two hits.The experiment was divided into control group(S),two-hits model group(HL),DEX group(HLD),and PDS group(HLP).The pathological changes of lung tissue were examined by HE staining.The expression of AQP1 was analyzed by RT-PCR,Western blotting and immunohistochemical staining.Results ① Significant inflammatory changes in pulmonary interstitial of rats in HL group were observed.However,in HLD group and HLP group,the pulmonary pathologic changes were much slighter.② AQP1 mRNA and protein expressions in lung tissues in HL group were significantly decreased compared with others groups(P

7.
National Journal of Andrology ; (12): 819-823, 2004.
Article in Chinese | WPRIM | ID: wpr-267806

ABSTRACT

<p><b>OBJECTIVE</b>To study the membrane mobility of aquaporin 7 (AQP7) by cloning stably transfected CHO cells with expression of pEGFP-C1-AQP7, in which AQP7 cDNA was fused downstream and in frame to pEGFP-C1 gene.</p><p><b>METHODS</b>The full sequence of AQP7 was amplified by RT-PCR and then recombined in the downstream of the green fluorescent protein gene in the pEGFP-C1 vector. The recombinant vector pEGFP-C1-AQP7 was stably transfected into CHO cells. With fluorescent microscopy, immunocytochemical stain and Western blot, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.</p><p><b>RESULTS</b>(1) The sequence of AQP7 cDNA of the Wistar rat was logged into the GenBank (access number: AY157737). (2) Identification demonstrated that pEGFP-C1-AQP7 fusion protein stably expressed in CHO cells. (3) With fluorescence microscopy, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.</p><p><b>CONCLUSION</b>The CHO cell line with stable pEGFP-C1-AQP7 expression was set up successfully for advanced research.</p>


Subject(s)
Animals , Aquaporins , Genetics , CHO Cells , Cricetinae , Cricetulus , DNA, Complementary , Genetics , Green Fluorescent Proteins , Genetics , Male , Molecular Sequence Data , Rats , Rats, Wistar , Recombinant Fusion Proteins , Reverse Transcriptase Polymerase Chain Reaction , Testis , Metabolism , Transfection
8.
National Journal of Andrology ; (12): 582-585, 2004.
Article in Chinese | WPRIM | ID: wpr-308296

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the use of free/total prostate-specific antigen ratio (fPSA/tPSA ratio) in improving the early diagnosis of prostate cancer.</p><p><b>METHODS</b>fPSA/tPSA ratio in the serum was analyzed prospectively in 187 men with tPSA ranging between 4.0 and 20.0 microg/L. All of them underwent ultrasound-guided sextant prostatic biopsies. Sensitivity, specificity, positive and negative predictive values were calculated by SPSS 10.0 software.</p><p><b>RESULTS</b>Prostate cancer detection rates were 18.1% and 22.5% when tPSA was within the ranges of 4.0-10.0 g/L and 10.0-20.0 g/L respectively. fPSA/tPSA ratio was more significant than tPSA in all the men. When the cut-off value of fPSA/tPSA ratio was set at 0.25, 90.5% and 87.5% of cancers could be detected; and 26.7% and 11.3% of biopsies could be avoided within the tPSA ranges of 4.0-10.0 g/L and 10.0-20.0 g/L, respectively.</p><p><b>CONCLUSION</b>The use of fPSA/tPSA ratio can improve prostate cancer detection and reduce unnecessary biopsies when tPSA is within the range of 4.0-10.0 microg/L and 10.0-20.0 microg/L.</p>


Subject(s)
Aged , Aged, 80 and over , Area Under Curve , Early Diagnosis , Humans , Male , Mass Screening , Middle Aged , Prospective Studies , Prostate-Specific Antigen , Blood , Prostatic Neoplasms , Diagnosis , Sensitivity and Specificity
9.
Article in Chinese | WPRIM | ID: wpr-410854

ABSTRACT

AIM: To find out whether different dosage of rare earth element-lanthanum can influence the expression of aquaporin 7(AQP 7) in the testis of rats. METHODS:Rats were fed with lanthanum nitrate [La(NO3)3] and killed 6 months later. Testes were then removed immediately to extract total RNA. Northern blot analysis is performed finally. RESULTS:0.1 mg/kg La(NO3)3 depressed the expression of AQP 7 in rat testis, while 20 mg/kg La(NO3)3 had no significant effect on it. CONCLUSION: AQP 7 expession is found in the rat testis; La(NO3)3 can depress the expression of AQP 7 in the rat testis.

10.
Article in Chinese | WPRIM | ID: wpr-578443

ABSTRACT

To study the underlying mechanism of vasovasotomy infertility, we set up the rabbit model and observed the levels of testosterone, IL-1 activity and TNFα, which represent the functions of monocytes and macrophages on the animal model of vasovasostomy, and probed into the in vitro effect of IL-1β on the production of testosterone by Leydig cells. Rabbit model was set up and divided into vasovasostomy fertility group (VFG) and vasovasostomy infertility group (VIG), and set up peer group of Vasectomy group (VG) and sham operation group (SOG) as controls. The results were as follows: The sperm density and total number is highest in SOG, with that in VFG markedly lower than that in SOG, but significantly higher than that in VIG.   1. VFG showed significant higher level of testosterone (Ts) than SOG, VG and VIG; Ts of VIG was significantly lower than SOG or VFG. In the vasovasostomy groups, sperm density is positively correlated to the level of testosterone.   2. The level of serum cortisol in VIG significantly higher than that in VFG, SOG, and VG. The level of serum cortisol negatively correlated to serum testosterone. 3. Serum estradiol(E2) was similar among all the groups (P>0.05). E2/T ratio is significantly different among the groups, with that in VIG highest and that in VFG lowest. 4. Serum IL-1 activity in VIG was significantly higher than that in SOG and VFG(P<0.05).   5. Serum TNFα and serum IL-1 activity share similar change, with TNFα in VG significantly higher than that in SOG(P<0.01), VIG was the highest among the four groups(P<0.01). Correlation test showed that serum IL-1 activity and TNFα level was negatively correlated to testosterone level; positively correlated to cortisol level and E2/T ratio. 6. Immuno-histo-chemistry of CD68 and TNFα in rabbits with vasovasostomy showed that in the testis tissue of SOG, interstitial tissue was normal with no filtration of inflammation cells including CD68 cells. In VFG and VIG, the tubules with resumed sperm production showed negative staining, and atrophic tubules and their surroundings were expression positive of CD68, with that markedly deeper in VIG than that in VFG. TNFα positive expressed granules was found only in atrophic convolted seminiferous tubules and their surroundings, with that deeper in VIG than that in VFG. 7. Culture Leydig cells were put in the following holes:(1) Control; (2) hCG; (3) IL-1; (4)IL-1 β+hCG; (5) IL-1 β+IL-1Ra; and (6) hCG+IL-1β+IL-1Ra. Determined the levels of testosterone by immuno-radiation technique. (1) IL-1 β showed no obvious effect on the Leydig cultured in vitro, but depressed the induction of Leydig cells by hCG to synthesize testosterone; (2) IL-1Ra blocked the depression of IL-1β Leydig cells' bio-synthesis of testosterone induced by hCG. In summary, one of the obvious changes in animals with vasovasotomy is low density of sperms, which is related to the lowering of testosterone. Low level of testosterone is related to the activation of monocytes and macrophages to produce IL-1 and TNFα, which negatively regulate the production of testosterone.

11.
Article in Chinese | WPRIM | ID: wpr-597675

ABSTRACT

AIM: Aquaporins(AQP) are very important for the water transport across cell membrane. Aquaporin 7(AQP 7) and aquaporin 8(AQP8) expressed in the rat testis, but the biophysical functions of these two aquporins in testis and the regulatory mechanism of their expression remain unclear. The aim of this study was to find out if the expression of these two aquporins was correlative with the function of testis, and if panaxadiol saponins (PDS) affected their expression. METHODS: 4 weeks, 8 weeks and 12 weeks old rats were divided into saline and PDS injection groups. Semi-quantified RT-PCR method was employed to detect aquaporin 7 and 8 gene expression: using total RNA extracted from rat testis as PCR template, then using optical scanner to measure the OD value of bands on agrose electrophoresis. OD value of target gene products was divided by which of contol gene. The ratio was identified as the quantity of target gene expression. Western blot was also employed to detect expression of these two proteins in the testis. RESULTS: ①OD value of AQP 7 mRNA products in testis of 4 weeks, 8 weeks, 12 weeks rats were 48 227, 51 536, 59 567 respectively and the ratio divided by OD value of control gene was 0.82, 0.85, 0.99; OD value of AQP 8 mRNA products in testis of 4 weeks, 8 weeks, 12 weeks rats were 23 092, 39 302, 43 316 respectively, the ratio divided by OD value of control gene was 0.39, 0.65, 0.72. Thus, AQP 7 and 8 mRNA expression increased with age. ②After PDS injection for 2 weeks. AQP 7 and 8 mRNA expression in 4 weeks old rats increased to 0.97 and 0. 72,which in control group were 0. 82 and 0. 39; the ratio decreased to 0.77 and 0.55 in 8 weeks old rats, which in control group were 0.85 and 0.65. There was no PCR product of neither aquaporins in 12 weeks old PDS injected rats, except products of control gene. ③ Western blot of AQP 7 and 8 protein showed little difference between PDS and saline injected 12 weeks old rats. CONCLUSION: ①Quantity of AQP 7 and AQP 8 expression was related to testis function of rats. ②PDS influenced the expression of AQP 7 and AQP 8 mRNA, perhaps by promoting the secretion of LH in pituitary.

12.
Article in Chinese | WPRIM | ID: wpr-592746

ABSTRACT

AIM: PSP94 has been shown promise as a potential prostate cancer marker and it was reported that PSP94 can inhibit the growth of prostate cancer cell in vitro and in vivo. This study aimed to construct recombinant human PSP94 expression vector. METHODS:The PSP94 cDNA was obtained from normal prostate tissue, and recombinant plasmid pUC19-PSP94 was constructed. The target gene was identified and sequenced. Then the PSP94 gene was inserted to the secretory expression vector. RESULTS:The gene sequence of PSP94 was identified. The recombinant vector was constructed. The secreted PSP94 was isolated and identified by Western blot. CONCLUSION:The recombinated PSP94 could expressed PSP94 successfully.

13.
Article in Chinese | WPRIM | ID: wpr-592700

ABSTRACT

AIM:Aquaporins (AQP) are very important for the water transport across cell membrane. There are at least 10 mammalian AQPs( aquaporins 0-9) distributed in various organs and different kinds of cells. Each AQP has a distinct organ distribution, and this distribution could be useful in presuming the biological function of the aquaporin. The aim of this study was to figure out the distribution of aquaporin 7 (AQP7) and aquaporin 8(AQP8).METHODS:Semi-quantified RT-PCR was employed in this research. The ratio of OD value of target gene products divided by which of control gene products was calculated. Among 35 organs, testis, epididymis, skin, muscle, rectum, lung, bronchus, lymph node, stomach, duodenum, jejunum, ileum, colon, pancreas, liver, gall bladder, spleen, mammary gland, uterus, placenta, tonsil, urinary bladder, thyroid came from normal area of removed samples during operation. cDNA library of Prostate, thymus, salivary gland, penis, carotiol artery, adrenal gland, occipital lobe of brain, temporal lobe of brain, frontal lobe of brain, parietal lobe of brain, mid brain, choroid plexus are purchased from OriGene biotechnique company.RESULTS:①AQP 7 mRNA was found in testis, muscle, gall bladder, carotiol artery, lymph node and adrenal gland, and maximum expression of AQP 7 was in testis.②AQP 8 mRNA was found in pancreas, testis, skin and colon. and maximum expression of AQP 8 was in pancreas.CONCLUSION:Coexistence of AQP 7 and 8 in testis was confirmed, which suggested that both of these two aquaporins were involved in the regulation of testis function.

14.
Article in Chinese | WPRIM | ID: wpr-531530

ABSTRACT

AIM: To establish the two-hit rat model with hemorrhage and lipopolysaccharide(LPS) and to study the effect of panaxadiol saponins(PDS) against acute lung injury.METHODS: Forty Wistar rats were divided randomly into 4 groups: sham operational group(S);two-hit groups with hemorrhagic shock-LPS(HL);dexamethasone pretreatment group(HLD) and PDS pretreatment group(HLP).The mean arterial blood pressure(MABP) was monitored dynamically by 4-channel physiological meter RM-6000,and pathological alteration of lung tissues was also observed.The levels of various serum enzymes,TNF-? and IL-6 were detected.RESULTS: MABP decreased in two-hit rat model with hemorrhage-LPS.The serum levels of aspartate aminotransferase,alanine aminotransferase,alkaline phosphatase,lactate dehydrogenase,creatine kinase,TNF-? and IL-6 increased significantly.Severe inflammatory change of pulmonary interstitium in HL group was also observed.CONCLUSION: Endotoxin injection following hemorrhage can be used to establish the animal model of acute lung injury.Similar with dexamethasone,PDS prevents lung tissue from seriously damage through increasing MABP and decreasing the level of serum enzymes,TNF-? and IL-6 levels.

15.
Article in Chinese | WPRIM | ID: wpr-529273

ABSTRACT

AIM:To study the effect of vitamin K3(VK3) on the induction of apoptosis in androgen-independent prostate cancer cell PC-3M in vitro.METHODS:Cell viability was estimated by MTT assay.AO/EB staining was performed to detect apoptotic cells.Apoptosis and the changes of cell cycle were detected by flow cytometry.NAC was used to observe the effect of growth inhibition by VK3.RT-PCR was used to confirm the changes in gene expression.Levels of intracellular peroxides were estimated by using an oxidation-sensitive fluorescent probe DCFH-DA.RESULTS:PC-3M cells growth was significantly inhibited by VK3(≥60 ?mol/L,P

16.
Article in Chinese | WPRIM | ID: wpr-525532

ABSTRACT

Aquaporins are small integral membrane proteins that selectively transport water and some small solutes in mammals, plants and lower organisms. Continued studies of the aquaporins are providing detailed tissue localization and the important physiological roles of aquaporins in urinary, respiratory, digestive and nervous systems. Based on the importance of aquaporin studies in the field of structural chemistry in biomembranes, American scientist Peter Agre won the 2003 Chemistry Nobel Prize for the discovery of water channels.

17.
Article in Chinese | WPRIM | ID: wpr-519511

ABSTRACT

AIM: To study the effects of 7 weeks prolonged haevy load swimming and Panaxadiol Saponins (PDS) on the expression of ?-actin in quadriceps muscle of thigh in rats. METHODS: After 7 weeks prolonged heavy load swimming training, the expression of ?-actin in quadriceps muscle of thigh was quantitatively examined by Northern blotting analysis in the condition with or without PDS treatment. RESULTS: The expression of ?-actin in rat quadriceps muscle of thigh in exercise+PDS group was significantly higher than that in sedentray control group and exercise+saline group at 24 th hour after intensive training. No statistical difference between exercise+androsan group and exercise+saline group was observed.CONCLUSION: PDS enhanced ?-actin expression in quadriceps muscle of thigh in rats.

18.
Article in Chinese | WPRIM | ID: wpr-519366

ABSTRACT

AIM: To explore the influence of ginsenoside Rh 2 on cultured PC-3M cell cycle in vitro . METHODS: DNA content was measured using -TdR incorporation assay. To analysis the cycle changes of PC-3M cells, flow cytometry was used in the experiment.RESULTS: The results indicated that the rate of -TdR incorporation was lower in Rh 2-treated cells than the control in a dose-dependent manner. Flow cytometry demonstrated that the pecent of G 1 phase treated with Rh 2 was higher than that of control. CONCLUSION: These results suggested that PC-3M cells cultured with Rh 2 for 24 h were blocked at G 1 phase, and Rh 2 inhibited the growth of PC-3M cells in vitro .

19.
Article in Chinese | WPRIM | ID: wpr-524633

ABSTRACT

0.05). Conclusions Silencing STAT3 gene can decrease STAT3 gene expressions , increase Bax gene expressions , induce cell apoptosis and suppress the tumor growth in the human breast carcinoma model by the RNAi technology .

20.
Article in Chinese | WPRIM | ID: wpr-522591

ABSTRACT

AIM: To investigate the protective effect and mechanisms of di-ao-xin-xue-kang (DK) on myocardial ischemia-reperfusion. METHODS: Models of myocardial ischemia-reperfusion were constructed with sixteen mongrel dogs. The left ventricular pressure (LVSP), the marximal/minimum rate of LVSP (?dp/dt max) and the serum aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA) were determined before and 90 minutes after occlusion, 120 and 240 minutes after reperfusion, and the MDA content in myocardial cell membrane prepared at 240 minutes after reperfusion was determined. The oxygen free radical was assayed with electron spin resonace spectroscopy (ESR) technique. RESULTS: ① LVSP and ?dp/dt max in the normal saline control (NS) group decreased with the time progress of occlusion-reperfusion, and it was the same in DK group, but the levels were significantly higher than that in NS group (P

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