Objective To discuss the best resistance factors of paclitaxel(Taxinol)on benign biliary scar fibrosis,in order to provide an effective basis for clinical prevention and treatment of benign biliary scar fibrosis.Methods Human bile duct epithelial cells were cultured in vitro,the prepared PTX at 0.001 uM,0.005 uM,0.1 uM,0.5 uMand 1 uMconcentration were separately added into cells for 48 h.The half inhibitory rate of BEC (IC50) were determined by MTT and the optimal concentration were confirmed.Human bile duct epithelial cells were cultured in 0 h,24 h,48 h and 72 h,the inhibitory rate of BEC at 100 nM,250 nM,and 500 nM PTX-Chitosan Sustained release membranes and the optimal concentration of PTX were determined by MTT and the optimal concentration of PTX-SRM were obtained.Human bile duct epithelial cells were cultured for 48 h and 72 h,the mRNA and protein expression ofα-SMA,E-cadherin,Vimentin were detected by Western Blot and Real-time PCR methods.Results The optimum resistance concentration of PTX to benign biliary scar was 250 nM.PTX and PTX-SRM could effectively inhibit the proliferation and transformation of BEC,and the best effective treatments to resist benign biliary scar fibrosis were low and middle concentrations of PTX-SRM,the best drug loading were 100 nMand 250 nM.The inhibition duration of PTX-SRMon BEC was longer than PTX alone(P<0.05).Conclusion The inhibition of PTX-SRMon BEC proliferation and transformation is better than the single drug of PTX,which provides a new scientific and feasible method for clinical prevention and treatment of benign biliary scar.

Objective To explore the effect of Paclitaxel-Chitosan Sustain film on growth, apoptosis and cell cycle of biliary fibroblasts cells. Methods Human biliary fibroblasts cells were cultured and treated with PTX-CSF and naked PTX,separately, untreated cells as blank control. The experiment was divided into five groups:untreated group, simple PTX-treated group (250nM) and low, medium and high chitosan sustained-release film PTX-treated group (100 nM, 250 nM, 500 nM). The proliferations of cells were determined by MTT assay. The apoptosis and cell cycle of cells were detected by FCM. Results The proliferation of biliary fibroblasts cells was inhibited by PTX-CSF with time-dependent and dose-dependent, and the inhibiting effect was more obvious than naked PTX treatment as the time went on. Meanwhile, PTX-CSF could inhibit the magration of bile duct fibroblasts induced by TGF-β1,and had longer effect than naked PTX. After 72 h, the apoptosis rate of cells treated with PTX-CSF was significantly higher than cells treated with naked PTX or untreated cells(P<0.05), the difference between naked PTX or untreated cells was not significant. Compared with untreated cells, the proportion of G 2/M in cells treated with PTX or PTX-CSF were significantly increased, and the former was sinificantly higher than the latter(P<0.05). Conclusion Compared with naked PTX, PTX-CSF has strong cytotoxic effects and obviously sustained-release effect. The effective concentration can be maintain for a long time by PTX-CSF, and it could be as the novel drug delivery system to continuously inhibit proliferation of bile duct fibroblasts.

Objective To investigate paclitaxel and ezrin-siRNA therapy for metastatic liver cancer.Methods Human metastatic liver cancer cells MHCC97-H were divided into four groups in vitro and in vivo,including control cells,exposure to paclitaxel alone,ezrin-siRNA alone,and the combination of paclitaxel and ezrin-siRNA.Nude mice lung lesions were formed by tail vein injection of tumor cells.The tumor cells'apoptosis rate,wound healing ability,the ability to cross an artificial membrane,and the numbers of metastatic lung lesions were measured.Results The apoptosis rate in normally cultured MHCC97-H tumor cells was 2.52％,compared to the other three groups at 11.66％,2.35％,7.38％ respectively.The wound healing ability was best in the negative control and weakest in cells treated with combination paclitaxel and ezrin-siRNA.The numbers of cells which could cross the artificial membrane in the negative,paclitaxel,ezrin-siRNA,and combination paclitaxel and ezrin-siRNA group were 52.5 ± 3.9,32.2 ± 3.2,26.6 ± 2.4,and 19.4 ± 1.1 respectively.The numbers of metastatic lung lesions in the nude mice (n =40) were 12.5 ±2.4,8.2 ± 1.5,7.0 ± 1.3,and 3.8 ± 0.3 respectively.Conclusions The ezrin-siRNA combined with paclitaxel therapy demonstrated a greater efficacy over single agent therapy for depressing liver cancer growth,motility,aggression,and metastasis in vitro and in vivo.

Background:It has been demonstrated that H + -K + -ATPase expression in human parietal cells had a tendency to increase with aging. However,the effect of aging on activity of H + -K + -ATPase is still unclear. Aims:To investigate effect of aging on activity of H + -K + -ATPase. Methods:Fifteen male Sprague-Dawley(SD)rats were divided into 4,21,24, 27 and 30 months group,and 19 healthy beagle dogs were divided into younger group,junior elderly group and senior elderly group. The activity of H + -K + -ATPase in gastric fundal mucosa was assessed. Results:The activity of H + -K + -ATPase in gastric fundal mucosa in 4,21,24,27 and 30 months old rats were(4. 850 ± 0. 312)μmol·mg - 1 ·h - 1 , (5. 466 ± 0. 379)μmol·mg - 1 ·h - 1 ,(6. 068 ± 0. 228)μmol·mg - 1 ·h - 1 ,(5. 733 ± 0. 767)μmol·mg - 1 ·h - 1 and (6. 223 ± 0. 428)μmol · mg - 1 · h - 1 ,respectively. With aging,H + -K + -ATPase activity in rats had a tendency to increase(F = 4. 519,P = 0. 031). The activity of H + -K + -ATPase in beagle dogs in younger group,junior elderly group and senior elderly group were(11. 087 ± 4. 320)μmol·mg - 1 ·h - 1 ,(8. 549 ± 3. 250)μmol·mg - 1 ·h - 1 ,(12. 071 ± 2. 820)μmol·mg - 1 ·h - 1 ,respectively. There was no statistically significant difference among the three groups(F =1. 339,P = 0. 290). Conclusions:With aging,the activity of H + -K + -ATPase in rats and beagle dogs does not decline, but even has a tendency to increase.

Objective To constructan eukaryotic expression recombinant plasmid named pEGFP-N2-PRNP .Methods Total RNA was extracted from alzheimer (AD) disease peripheral blood ,and the PRNP gene was amplified by reverse transcription-poly-merase chain reaction(RT-PCR) .By using gene recombination technique ,human PRNP cDNA was inserted into retroviral vector pEGFP-N2 .The recombinant plasmid was identified by a pair of specified primers containing the restriction sites of Xho Ⅰ and EcoRⅠ .Results The PRNP gene could be obtained by RT-PCR ,the recombinant plasmid was identified by restriction endonucle-ase analysis ,PCR and sequence analysis ,and the expression vector pEGFP-N2-PRNP ,which could be stably expressed in SH-SY5Y cells .Conclusion The recombinant plasmid pEGFP-N2-PRNP is constructed successfully ,Which offers a basic for the further re-search on PRNP biological fuction .

Recent studies showed that gastric mucosa was more susceptible to injury by invasion factors with aging, however,the studies were mainly on gastric antral mucosa,fundic mucosa was rarely studied.Aims:To investigate the effect of aging on related biological activity factors in gastric fundic mucosa in beagle dogs.Methods:Nineteen beagle dogs were assigned into younger group (aged 1-5 years),junior elderly group (aged 6-8 years)and senior elderly group (aged≥9 years).The contents of MDA,LPO,MPO in gastric fundic mucosa were determined by TBA method.The contents of PTEN,TE,survivin,caspase-3,caspase-9,ZO-1,CGRP,VEGF,COX-1 and COX-2 were assessed by ELISA.Results:Compared with younger group and junior elderly group,contents of MDA,LPO,MPO,PTEN,TE,ZO-1 ,CGRP,VEGF, COX-1 and COX-2 were significantly increased in senior elderly group (P<0.05),no significant differences in contents of caspase-3 and caspase-9 were found among the three groups (P>0.05 ).The content of survivin in junior elderly group and senior elderly group was significantly decreased when compared with younger group (P =0.000 ).Conclusions:Disadvantaging changes of biological activity factors are found in gastric fundic mucosa in elderly beagles dogs,however, gastric mucosal blood flow, mucosa regeneration and epithelial tight junction related biological activity factors are significantly increased in senior elderly beagle dogs,which may be a phenomenon of degeneration-compensation.

OBJECTIVE: The purpose of this study is to assess the effects of aging on air-conducted sound elicited cervical vestibular evoked myogenic potential (cVEMP) and ocular vestibular evoked myogenic potential(oVEMP) in normal people. METHOD: Sixty normal subjects were recruited for the vestibular evoked myogenic potential examination. Among them, 20-40 year-old group was setted as the first group, 41-60 year-old group was setted as the second group, > 60-year-old group was setted as the third group. Each group included 20 normal subjects. SPSS 16.0 software was used to analyze the data. RESULT: As the age growing, P1 latency prolonged, N1-P1 amplitude decreased, and amplitude asymmetric ratio increased in cVEMP; N1 latency prolonged, P1-N1 amplitude decreased, and amplitude asymmetric ratio increased in oVEMP. CONCLUSION With the age growing, the time of otolithic organ input pathways prolonged, the function of otolithic organ decreased and the decrease level may be asymeetrical on both sides.
Adult ; Aging ; Eye ; Head ; Hearing ; Humans ; Middle Aged ; Otolithic Membrane ; Vestibular Evoked Myogenic Potentials ; Vestibule, Labyrinth ; Young Adult

Objective To investigate the effect of aging on gastric pepsinogen secretion through observing histological changes of gastric fundal mucosa and ultrastructure of gastric chief cells in human and Beagle dogs at different ages.Methods Fifty middle-aged and elderly indigestion patients with gastroscopy were selected as study objects and divided into young and middle age group (age 20-59 years,n=19),junior elderly group (age 60-69 years,n=11),middle elderly group (age 70-79 years,n=10),and senior elderly group (no less than 80 years,n=10).In addition,nineteen healthy Beagle dogs were also selected as study objects and divided into young and middle age group (age between 1 to 5 years,n=8),junior elderly group (age six to eight years,n=5),and senior elderly group (no less than 9 years,n=6).The histology and morphology of gastric fundal mucosa of human and Beagle dogs were observed under light microscope.Then the thickness of lamina propria was measured and the number of gastric chief cells was counted.The ultrastructure of gastric chief cells was observed under electron microscope,and the area percentages of secretory granule(also called mucous granule)area in cytoplasm of chief cell was calculated.The data was analyzed by one-way analysis of variance.Results No significant histological and morphological changes of gastric fundal mucosa were observed in human and Beagle dogs at different ages.There was no significant difference in the thickness of lamina propria among different ages in human and Beagle dogs (all P＞0.05).The number of human chief cells of young and middle age group,junior elderly group,middle elderly group and senior elderly group was 71.79±16.85,52.73±16.60,57.10±20.21 and 43.70 ±16.89/high power field (HPF),respectively,which decreased as age increased (F=6.431,P=0.001).The numbers of chief cells of young and middle age group,junior elderly group,and senior elderly group of Beagle dogs were 328.38 ±32.36,341.20 ±42.49 and 225.67 ± 52.19/HPF,respectively,which decreased as age increased (F=13.647,P＜0.01).Aging-related degeneration was founded in ultrastructure of gastric chief cells in human and Beagle dogs including rough endoplasmic reticulum dilation,ribosones missing,secretory granule decreasing and so on.The area percentages of secretory granule area in cytoplasm of human chief cells of young and middle age group,junior elderly group,middle elderly group and senior elderly group were (67.28±3.79)％,(66.88±4.84)％,(65.63±7.10)％ and (56.25±8.47)％,respectively,which decreased as age increased (F=6.069,P=0.002).The area percentages of secretory granule area in cytoplasm of Beagle dogs' chief cells of young and middle age group,junior elderly group,and senior elderly group were (58.83 ± 3.07) ％,(52.34 ± 4.73) ％ and (39.89 ± 4.46) ％,respectively,which decreased as age increased (F=38.837,P＜0.01).Condusion As age increased,the ability of pepsinogen secretion of fundal gland decreased,which may be one of the reasons of functional dyspepsia in the eldly.

Background:It has been demonstrated that acid secretion-related gastric fundic structures do not degenerate in elderly individuals. As specimens obtained from human body are greatly influenced by environmental factors,the results need to be further verified by standard animals. Aims:To investigate the aging-related changes of histology of gastric fundus and ultrastructure of fundic gland in beagle dogs. Methods:Nineteen healthy beagle dogs were divided into three age groups,8 in younger group(aged 1-5 years),5 in junior elderly group(aged 6-8 years)and 6 in senior elderly group (aged ≥9 years). Gastric specimens obtained from the fundus were examined by light microscope,the thickness of lamina propria and percentage of parenchymal area of fundic gland in lamina propria were measured,and the parietal cells,chief cells and endocrine cells were counted. Ultrastructure of fundic gland cells were observed by electron microscopy,and the area percentages of related organelles were calculated. Results:The thickness of lamina propria in gastric fundus of beagle dogs showed a decline tendency with aging,but no significant differences were found among the three age groups(P >0. 05);in senior elderly group,a marked decrease in density of fundic gland was observed and with an increase of interstitial tissues. The amount of parietal cells and the area percentages of mitochondria and tubulovesicle in parietal cells showed no aging-related reduction(P > 0. 05),while the amount of chief cells,area percentages of zymogen granule in chief cells,and mucous granule in mucous neck cells were significantly decreased in senior elderly group(P < 0. 05). The amount of endocrine cells increased gradually with aging(P < 0. 05). Conclusions:Aging-related degeneration is not existed in acid secretion-related parietal cells and organelles in gastric fundus of beagle dogs. Furthermore,endocrine cells which secret histamine,gastrin,and somatostatin,etc. for stimulating acid secretion even increased. These changes might be a compensatory mechanism for acid secretion in elderly individuals.

Objective To investigate the effects of intervention with the fluoxetine and the enriched environment on chronic stress induced depression behavior of rats,and the changes of myelin basic protein in hippocampus and prefrontal regions.Methods 50 adult male SD rats were randomly divided into control group,fluoxetine group,model group,enriched environment (EE) group and EE plus fluoxetine group.Fluoxetine group,model group,EE group and EE plus fluoxetine group underwent chronic unpredictable stress stimulus in the first to third week,and fluoxetine group,EE group,EE plus fluoxetine group underwent the intervention with EE and (or) fluoxetine in the fourth to sixth week.The changes of behavior in rats were evaluated by sucrose water consumption,open field test and weight changes.The content of MBP in each subregion of hippocampus and prefrontal regions of rats was measured with immunocytochemical methods.Results At the third weekend,the assessed behaviors of stressed rats decreased significantly compared with control group (P＜0.05);and at the sixth weekend,the behaviors of stressed rats restored after treated with EE and (or) fluoxetine.The content of MBP in the rat hippocampus CA1,DG area and prefrontal area of model group declined clearly compared with control group (mean density of model group orderly:0.199±0.024,0.204±0.021,0.225±0.028;control group orderly:0.279±0.034,0.288±0.043,0.308±0.053,P＜0.05).The content of MBP in the rat of fluoxetine group,EE group and EE plus fluoxetine group increased obviously compared with model group (fluoxetine group orderly:0.259± 0.047,0.266± 0.052,0.284 ± 0.031;EE group orderly:0.257±0.038,0.258±0.042,0.286±0.037;EE plus fluoxetine group orderly:0.271± 0.046,0.279±0.040,0.289±0.041,P＜0.05).Conclusion The depression-like behavior of rats induced by chronic unpredictable stress is associated with the change of the content of MBP in hippocampal CA1,DG area and prefrontal area;and the depression-like behavior and the content of MBP decrease are reversed after the intervention with fluoxetine and EE.