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.

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.

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).

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.

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.

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.

Objective:Based on the microarray data mining method, the function and pathway of differential genes were analyzed after the differential genes were screened. At the same time, the core genes that determine the prognosis of pediatric hepatoblastoma were screened by coexpression network, and their predictive ability was evaluated.Methods:The microarray expression profile of pediatric hepatoblastoma used in this study was from the European Institute of bioinformatics (http: //www.ebi.ac.uk/embl/). The deadline for data collection was December 31, 2018. Firstly, the differentially expressed genes (gene expression level increased to 2 times or decreased to 1/2 of the original) were screened by SAM method, then the core genes were screened by coexpression network model based on dimension reduction principle, and the gene regulation evaluation score was calculated by MCODE algorithm to evaluate its regulation ability in the whole network model.Results:According to the enrichment results of 213 differentially expressed genes, the highest enrichment degree of signal pathway was metabolic pathways (2 122.529). The misjudgment rate of signal pathway enrichment results was less than 0.001, and the misjudgment rate was statistically significant by SAM method ( P<0.001). A total of 213 differentially expressed genes in different prognosis groups were used as the basis for the construction of the coexpression network. A total of 12 differentially expressed genes were included in the coexpression network. Using the poor prognosis group as the experimental group, and the better prognosis group as the control group, the MCODE algorithm was used to calculate the gene regulatory ability score. The results showed that the highest gene for determining the prognosis control ability of children hepatblastoma was ADH1A gene with a score of 19. In addition, the regulatory ability scores of HAO1, ADH1B, ALDOB and DPYS genes were higher than or close to 5, so they could be considered as the core genes in the coexpression network module. Conclusions:According to the results of coexpression network model, ADH1A gene is closely related to the occurrence and development of hepatoblastoma in children, and its molecular biological evidence needs to be further explored to guide the clinical development of tumor targeted intervention therapy.

Aim To investigate the effect of MTX(included(±)MTX,(+)MTX and(-)MTX)on the proliferation of ECV304 cells and to explore its mechanisms.Methods ECV304 cells were cultured.The cell proliferation was determined by MTT.The morphological changes were inspected by inverted microscope.Cell cycle phases were assayed by propidium iodide staining flow cytometry.Results ECV304 cells were treated with(+)MTX,(-)MTX and(±)MTX at 1～150 μmol·L~(-1) for 24,48,72 h.The results showed that the proliferation of ECV304 cells was significantly inhibited under different conditions.The order of the inhibited efficacy was(+)MTX>(±)MTX>(-)MTX.The morphology of ECV304 cells were changed by(+)MTX,(-)MTX and(±)MTX treatment,which included the cell shrinkage,chromatin condensation.After administration of 10 μmol·L~(-1) of(+)MTX,(-)MTX and(±)MTX for 48 h,the cell cycle phases were assayed by propidium iodide staining flow cytometry.The result showed DNA replication was interfered by(+)MTX,(-)MTX and(±)MTX treatment.Conclusions The proliferation of ECV304 cells has the chiral selective effects by(+)MTX and(-)MTX treatment,and the inhibition on ECV304 cells proliferation of(+)MTX is significantly stronger than that of (-)MTX.

BACKGROUND: Salvia miltiorrhiza is widely used to treat angina cordis, ischemic stroke and other ischemic cardiovascular diseases. However, the effects of Salvia miltiorrhiza on ovariectomized rats remain unclear.OBJECTIVE: To observe the effects of Salvia miltiorrhiza on the body mass, food intake, and levels of blood lipids and malondialdehyde (MDA) in ovariectomized rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University.MATERIALS: The experiment was performed in the Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province and the laboratory of Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University from November 2005 to December 2006. Twenty-four healthy female SD rats of 3 months old and (220±2) g were selected. Salvia miltiorrhiza water decoction (equal to 1 g/mL crude drug) was identified and extracted by Drug Control Institute of Gansu Province; MDA kit was purchased from Nanjing Jiancheng Institute of Bioengineering.METHODS: ①The rats were randomly divided into three groups with 8 rats in each group: sham-operated group,ovariectomized group and Salvia miltiorrhiza group. The rats were underwent a bilateral ovariectomy except those in the sham-operated group, which were subjected to a removal of bilateral fat as much as ovariectomized group with the ovaries remained. Rats in sham-operated group and ovariectomized group freely drank water; rats in Salvia miltiorrhiza group freely took 1% water extracts from Salvia miltiorrhiza postoperatively, and the concentration of Salvia miltiorrhiza gradually increased to 12% on the eighth day, which was lasted until the end of the experiment (55 days). ②The food intake of rats in each group was monitored daily, and the body mass was measured every five days. At the end of the experiment, femoral artery blood samples of rats were collected to determine the levels of blood lipids. At the same time,MDA was measured according to the kit.MAIN OUTCOME MEASURES: The body mass, food intake, levels of blood lipids and malondialdehyde in each group.RESULTS: Twenty-four rats all entered the result analysis. ①The body mass of rats in 3 groups was nearly the same before operation (P ＞ 0.05). While the body mass in ovariectomized group on the postoperatively 10th, 20th, 25th, and 55th days was significantly higher than those in sham-operated group (P ＜ 0.01). The body mass in Salvia miltiorrhiza group on the postoperatively 20th, 25th, and 55th days was significantly lower than those in ovariectomized group (P ＜0.05-0.01). ②The food intake in ovariectomized group on the postoperatively 15th, 40th, and 55th days was significantly higher than those in sham-operated group (P ＜ 0.05-0.01), and that in Salvia miltiorrhiza group was significantly lower than those in ovariectomized group at those 3 time points (P ＜ 0.05-0.01). ③At the end of the experiment, the levels of total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol and triglyceride in ovariectomized group were significantly higher than those in sham-operated group (P ＜ 0.05-0.01). The levels of triglyceride and MDA in Salvia miltiorrhiza group were significantly lower than those in ovariectomized group (P＜0.01, 0.05).CONCLUSION: Salvia miltiorrhiza can significantly reduce the body mass and levels of triglyceride and MDA in ovariectomized rats.