1.Rab13 regulates the distribution of occludin and F-actin at the blood-testis barrier via PKA activity in rats
Wenhui SU ; Xiaoyu JIA ; Xiaona MENG
Acta Laboratorium Animalis Scientia Sinica 2015;(4):359-364
Objective To investigate the regulation of blood-testis barrier by Rab13-PKA pathway in rats.Meth-od First, shRNA vector targeting at Rab13 was constructed and then the Rab13 shRNA was transfected into the rat testis by injection.Western blot was used to detect the knock-down effect of Rab13 and the expression of blood-testis barrier ( BTB) constituent proteins.PKA activity was detected by autoradiography and scintillation counting.Further, immunoflu-orescence analysis and phalloidin staining were applied to observe the distribution of occludin and F-actin, respectively. Results The expression level of Rab13 in the testis was reduced by approximately 70%after transfection of Rab13 shRNA as compared with the non-targeted control group ( P<0.01 ) , while the expression of BTB constituent proteins remained unchanged.PKA activity was significantly increased after Rab13 RNAi transfection (P<0.01).The distribution of occlu-din at BTB was remarkably increased after Rab13 RNAi silencing around stage VIII but not at other stages of the seminifer-ous epithelial cycle.The assembly of F-actin at BTB was also intensified in Rab13-silenced testis.Both the changes of dis-tribution of occludin and F-actin induced by Rab13 shRNA were found to be antagonized by the PKA specific inhibitor H89.Conclusions Rab13 can modulate the distribution of occludin and F-actin at the blood-testis barrier in rats by regu-lating PKA activity, which may participate in the regulation of BTB function.
3.The clinical efficacy ofNingmitai capsule combined with tolterodine in the treatment of overactiver bladder after transurethral resection of the prostate
Min JIA ; Xiaoyu ZHANG ; Shuo LIU ; Shengping HU
International Journal of Traditional Chinese Medicine 2015;(8):695-698
Objective To observe the clinical efficacy of Ningmitai capsule combined with Tolterodine in the treatment of overactiver bladder after transurethral resection of the prostate(TURP).Methods 63 TURP patients with bladder disease (OAB) in our hospital from October 2012 to October 2012 were selected and randomly divided into a treatment group (n=32) and a control group (n=31). From the date of operation, the control group was treated with tolterodine 2 mg, 2 times a day, while the treatment group was additional treated with Ningmitai capsule 4mg, 3 times a day based on the control group. All patients stopped to take medicine 5 days after catheter removal. The pain frequency and duration of bladder spasm after TURP in catheterization period and the urine volume per time, the number of urgent incontinence, and the number of urgent micturition average day in automatic micturition period were scored by OABSS.Results The number of bladder spasm in the first postoperative day(3.5 ± 0.5vs.4.4 ± 0.8,t=2.650), the second day(1.5 ± 0.9vs.1.8 ± 0.2,t=2.350) and the third day (0.4 ± 1.6vs. 1.1 ± 1.8,t=2.210) of the treatment group were all less than the control group (P>0.05). The 24 h average frequency of urination after catheter removal (6.2 ± 1.3vs. 9.4 ± 1.8,t=2.710), the average number of nocturia (1.5 ± 0.4vs. 3.9 ± 1.0,t=2.580), the average number of 24h urinary urgency (1.1 ± 0.3vs. 3.2 ± 0.8,t=2.660), the average number of incontinence in 24 h (0.5 ± 0.2vs. 2.4 ± 0.6,t=2.700) and OABSS total score (4.6 ± 1.2vs. 6.9 ± 2.1,t=2.470) of the treatment group were all better than the control group (P<0.05). Conclusion Ningmitai capsule combined with tolterodine in the treatment of overactiver bladder after TURP has significant clinical effect, helping patients recovery and improving quality of life.
4.Effects on the pharmacokinetics of furosemide after acute exposure to high altitude at 4010 meters in rats.
Wenbin LI ; Rong WANG ; Hua XIE ; Juanhong ZHANG ; Xihui XIE ; Xiaoyu WU ; Zhengping JIA
Acta Pharmaceutica Sinica 2012;47(12):1718-21
The paper is to report the pharmacokinetics of furosemide in rats living at plain area and high altitude. After intragastric administration of furosemide (2.87 mg x kg(-1)), serial blood samples (0.5 mL) were collected by retro-orbital puncture at 0, 20 min, 40 min, 1, 1.5, 2, 3, 4, 6, 8, 12 and 24 h, samples were determined by LC-MS/MS, and plasma concentration-time data were analyzed by DAS 2.0 software to get the related pharmacokinetic parameters. The main pharmacokinetic parameters: area under curve (AUC), mean residence time (MRT), the biological half-life (t1/2) and the peak concentration (C(max)) of furosemide, were significantly increased at high altitude, the time to reach peak concentration (t(max)) and clearance (CL) was significantly decreased. This study found significant changes on the pharmacokinetics of furosemide under the special environment of high altitude. This finding may provide some references for clinical rational application of furosemide at high altitude.
5.Establishment and preliminary application of the McA-RH7777/Buffalo hepatoma rat model with bioluminescence imaging
Lichao PAN ; Aiqun ZHANG ; Jia BAI ; Xiaoyu TAN ; Xinlan GE ; Yanhua LAI ; Jiahong DONG
Chinese Journal of Hepatobiliary Surgery 2015;21(5):332-336
Objective To establish a luciferase labeled McA-RH7777 hepatoma rat model,which could be used for gross observation to further observe the effect of selective ligation of the portal vein and bile duct on tumor growth and metastasis.Methods The luciferase gene was transfected into rat McA-RH7777 hepatoma cells with pCDH-puromycin-CMV as the carrier,which were subcutaneously inoculated into Buffalo rats.Tumor pieces were then heterotransplanted into the left lateral lobe of the allogenic rat liver to observe the tumor growth in vivo.After the successful hepatoma modeling,the rats were randomly divided into three groups,namely the implanted portal vein group with combined portal vein and bile duct ligation,the implanted portal vein group with single portal vein ligation and sham operation group.The rats were executed at the 1 st week and 2nd week after ligation,and the livers were dissected to record the tumor growth and metastasis inside and outside the liver,respectively.Results The tumor formation rates of Buffalo rats after subcutaneous and intrahepatic implantation were both 100%.The fluorescence signal implanted into the liver lobe could be observed in vivo after the intrahepatic implantation of luciferase transfected Luc-McA-RH7777 at 2nd week,the range and intensity of which increased over time.Only local tumor growth could be found at the 4th week,without obvious intrahepatic and lung metastasis.However,both an increased in situ tumor volume and the pulmonary metastasis could be observed in the implanted portal vein group with combined portal vein and bile duct ligation at 2nd week after the ligation.Immunohistochemistry showed AFP positive immunoreactions in the vast majority of intrahepatic tumor cells and Luc positive immunoreactions in part of tumor cells.Conclusion Luc-McA-RH7777 cells could be used to establish the heptoma rat model and the in vivo analysis within the Buffalo rat liver demonstrated that the combined ligation of the portal vein and bile duct can accelerate the development and metastasis of liver cancer.
6.Need-based design of medical mobile learning platform
Juanping WU ; Wenjie LI ; Xiaoyu WANG ; Wen ZHANG ; Jia XUE ; Peifeng HE ; Jianguang WU
Chinese Journal of Medical Library and Information Science 2014;(2):68-71
The medical mobile learning platform was constructed according to the information need of teachers and students in Shanxi Medical University, Changzhi Medical College, and Fenyang Medical College.The teaching and learning resources in Shanxi Medical University were integrated into the platform which could thus provide a variety of interactive learning ways for its users and users could rapidly find out their interested information resources. However, the platform construction needs the implementation of incentive measures, and regulations and rules for the protection of intellectual property rights.
7.Establishment and application of a high-throughput screening assay for premature activation of HIV-1 precursors.
Quan ZHANG ; Xiaoyu LI ; Zhenlong LIU ; Pingping JIA ; Xiaolu WEI ; Lixun ZHAO ; Jiandong JIANG ; Shan CEN
Acta Pharmaceutica Sinica 2010;45(2):247-52
Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).
8.Establishment and application of a screening anti-HIV-1 drug model targeted nuclear trafficking of virus RNA.
Zhenlong LIU ; Xiaoyu LI ; Quan ZHANG ; Pingping JIA ; Liang YANG ; Xiaolu WEI ; Jiandong JIANG ; Shan CEN
Acta Pharmaceutica Sinica 2010;45(2):257-62
The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway. Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly, reveals a target for anti-HIV drug development. In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA nuclear export. This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner. Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP. The Z' score of this model is 0.8220. Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition. IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.
9.The impacts of white matter ischemic lesions in different regions on cognitive domains
Xiaoyu JIA ; Aijuan ZHANG ; Qi WANG ; Shaobo WANG ; Hongcai WANG ; Yanping XIN ; Jing CHEN
Chinese Journal of Geriatrics 2012;31(3):211-214
Objective To study the impacts of white matter ischemic lesions in various regions on the different cognitive domains of the patients. Methods 120 patients with white matter ischemic lesions were divided into subcortex,semi oval center,peri-ventricle,mixed regions according to MRI imaging (n=30 cases for each group). The 30 healthy control subjects were enrolled.Cognitive functions were evaluated by mini mental status scale (MMSE), montreal cognitive assessment (MOCA),object memory test (FOM),rapid verbal retrieve (RVR),block design (BD)and digit span (DS). Results The score of language in subcortical group (0.36±0.49) was lower than in control group (1.00±0.00) (P=0.011).There was no significant difference in RVR scores between mixed regions group and peri- ventricle group [(27.00 ± 9.22) vs. (32.30 ± 7.78) P =0.067],while RVR scores in mixed regions group (27.00± 9.22) were increased as compared with subcortex,semi oval center and control groups [(38.21±11.93),(35.94=9.53) and (37.00±3.16),respectively] (F=3.462,P=0.013).There was no difference,in BD scores between mixed regions group and semi- oval center group [(21.20± 9.21) vs.(25.63±12.10).P 0.070] but the mixed group scores were decreased as compared with subcortex, peri-ventricle, control groups [(37.14±10.43),(36.80± 14.27),(40.30±6.29),F=7.795,P=0.000].The scores of immediate verbal memory,calculation,short-term memory,visual spatial ability and executive were reduced in mixed regions group than in other groups (P=0.034,0.030,0.016,0.000).There was no difference in orientation score in MOCA and MMSE among the groups (P=0.256 and P=0.325).Conclusions Ischemic white matter lesions may lead to cognitive impairments depending on different region lesions. The obvious impact of peri-ventricle lesion is on memory, subcortex lesion on language,semi-oval center lesion on recognition and construction of images,while wide range of cognitive impairment may be attributed to the lesion in mixed regions.The scale of the MOCA is helpful and sensitive for identifying the presence of early cognitive impairment.
10.Immunogenicity of tandem expression recombinant EG95s protein of Echinococcus g ranulosus
Hong JIA ; Weifeng YUAN ; Jie LI ; Shaohua HOU ; Xiaoyu GUO ; Ting XIN ; Shichun MA ; Hongfei ZHU
Chinese Journal of Zoonoses 2014;(8):843-847
In order to analyze the immunogenicity of the recombinant EG95s protein ,the recombinant plasmids of pET-1EG95s ,pET-2EG95s and pET-3EG95s which containing respectively 1 ,2 ,and 3 copies EG95s were induced to express HIS-1EG95s ,HIS-2EG95s and HIS-3EG95s ,and then the proteins were purified and identified by western-blotting .The same im-mune process was used ,and 8 weeks-old BALB/c mice were immunized ,then its immunogenicity was analyzed by detecting an-tibody levels in mice by indirect ELISA method .Results showed that for recombinant EG95s proteins after transformation , HIS-1EG95s ,HIS-2EG95s ,and HIS-3EG95s also retained immunogenicity and could induce specific antibodies in mice .One week's late after the first immunization with HIS-1EG95s ,the antibody level of was significantly higher than HIS-2EG95s and HIS-3EG95s .But began from 2 weeks after immunization ,the antibody level of HIS-3EG95s was always higher than that of HIS-1EG95s group during the period of the immune .Both the final antibody titers after immunization of HIS-1EG95s and HIS-2EG95s groups was 1∶819 200 ,while HIS-3EG95s group was 1∶163 840 0 .HIS-1EG95s ,HIS-2EG95s and HIS-3EG95s all induced IFN-γin immune mice ,but the difference was not significant .The HIS-1EG95s showed lower response to Echinococ-cus granulosus positive serum than HIS-2EG95s and HIS-3EG95s .It’s indicated that the HIS-1EG95s and HIS-3EG95s also had good immunogenicity .HIS-3EG95s make recombinant protein immunic effects more lasting ,and benefit to generate more long-lasting protective immunity .This study provides the scientific basis for the immunization of echinococcosis (hydatidosis) .