Nucleic acid aptamers are ligands with high specificity and affinity for their targets, which are screened from large oligonucleotide pools by the SELEX technology. Their characteristics are superior to antibodies in many aspects, which make them important in the research of recognition of molecules. They can be used to determine the levels and inhibit the bioactivity of their targets. There're broad application prospects for aptamers in the diagnosis and treatment of tumor.

Adult ; Choanal Atresia ; etiology ; therapy ; Constriction, Pathologic ; etiology ; Humans ; Male ; Nasopharyngeal Diseases ; etiology ; therapy ; Otorhinolaryngologic Surgical Procedures ; adverse effects ; Palate ; surgery ; Postoperative Complications ; therapy ; Uvula ; surgery

Objective To study the effect of diabetes on spatial memory ability and spatial associative memory ability in rats. Methods 70 SD rats( 180±20 g) were randomly divided into 3 groups: control group,type1 diabetes group and type2 diabetes group. Type1 and type2 diabetic rat models were set up by streptozocin (STZ) intraperitoneal injection and high fat forage raise. The blood glucose was determined. After rat diabetic model established 1 month and 3 months, respectively, the Morris water maze experiments were implemented,including 4 days' place swimming and 1 day's space exploration. Results After 1 month, diabetic rats' spatial memory ability and spatial associative memory ability were not affected. After 3 months, in place swimming test,the escape latency of two diabetic rat groups was obviously longer than that of the control group (P＜0.05). From the second day of the experiment, escape latency of the control descended sharply, while that of the diabetic group descended slowly. There was no difference between type 1 and type2 diabetic groups in escape latency (P＞0.05). After 3 months, in the space exploration test, when rats were put into the maze from I quadrant which already trained, swimming time in the platform quadrant was shorter, the other parameter scores were lower of the two diabetic model groups, contrast to the control group (P＜0.05). The parameter scores of type2 diabetic group were lower slightly than type1 diabetic group. When rats were put into the maze from IV quadrant for which never trained, the parameter scores of two diabetic groups were lower than that of the control group (P＜0.05) and the total score of type 1 group was lower than that of type2 group. Conclusion The spatial memory ability and spatial associative memory ability of type 1 and type2 diabetic rats descended. In the experiment, the spatial memory ability of type2 diabetic rats was more significantly affected than that of type1 diabetic rats. By contrast,type 1 diabetic rats' spatial associative memory ability descended greatly than that of type2 diabetic rats.

Objective To study the effects of diabetes on hippocampal synaptic plasticity in perforant path-dentate gyrus pathway (PP-DG) in rats. Methods 70 SD rats( 180±20) g were divided into 3 groups at random: control group, type1 diabetes group (DM1)and type2 diabetes group (DM2). After Morris water maze test, 15 rats that showed worse spatial memory ability were selected in each model group to investigate the variation of paired-pulse facilitation (PPF) and the range of synaptic plasticity. Field potentials were recorded in the dentate gyrus of the dorsal hippocampus by stimulating the perforant path. Results Contrast to the control group, diabetic rats' hippocampal LTP were depressed (P<0.05), and type1 diabetic rats' LTP reduced much more. Diabetic rats' PPF ratio was reduced contrast to the control group (P<0.05). Conclusion Type1 and type2 diabetes impaired synaptic plasticity of hippocampal PP-DG pathway in rats, which conformed the results of water maze test.

China ; Congresses as Topic ; Humans ; Pharyngeal Diseases ; Tracheal Diseases

Objective To evaluate the inhibitory effects on Candida albicans of vagina cells transferred with antimicrobial peptide LL-37 and human defensin 5 (HD5) recombinant plasmids and observe secretion of IL-8.Methods ( 1 ) The epithelial cells from female vagina were culture primarily.The pcDNA3.1 ( + )/HD5-EGFP and pcDNA3.1 ( + )/LL-37-EGFP eukaryotic recombinant plasmids were separately or jointly transferred into the fourth generation of vaginal epithelial cells.Two test groups were defined:one test group was no Candida albicans group including four subgroups which were untransferred group,HD5 group transferred with pcDNA3.1 ( + )/HD5-EGFP,LL-37 group transferred with pcDNA3.1 (+)/LL-37-EGFP,combining transferring group transferred with pcDNA3.1 ( + )/HD5-EGFP and pcDNA3.1 ( + )/LL-37-EGFP; the other test group was with Candida albicans group which the Candida albicans were coincubated with the four subgroups described above.(2) For examination of cytokines and chemokines,at 6,12,24 and 48 hours,the supernatant of every group was collected.ELISA was applied to detect the levels of LL-37,HD5 and IL-8.At each time point,the growth inhibition of Candida albicans was calculated by glucose consumption testing.Results ( 1 ) The max level of LL-37,HD5 and IL-8 reached max level after being transferred for 24 hours,then showed decreasing trend.The secretion of LL-37,HD5 and IL-8 was significant higher in combining transferring group in Candida albicans group than other groups,and the secretion level of LL-37,HD5 and IL-8 was (100.16 ±0.81 ) ng/ml,(58.50 ±2.08) μg/ml and ( 101.03 ± 1.59) pg/ml (P ＜0.01 ).(2) In different time point,the absorbance of each subgroup without Candida albicans declined slowly,and there were no statistically significant difference (P ＞0.05 ),as while as in LL-37 subgroup and HD5 subgroup with Candida albicans.In group with Candida albicans,the absorbance of combining transferring subgroup were 3.210 ± 0.010,3.150 ± 0.030,3.099 ± 0.030 and 2.970 ±0.040 at 6,12,24 and 48 hours,respectively,which was significantly higher than those in the other cells (P ＜ 0.01 ),and the declined trend was the slowest.Conclusions The antifungal ability of vaginal epithelial cell became stronger after being transferred with LL-37 and HD5 recombinant plasmids.LL-37 and HD5 could also possess immunomodulatory activity and induce chemokine IL-8 production.

Objective This study is to investigate cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway gene expression pattern of peripheral blood mononuclear cell(PBMC) of primary sjgren syndrome patients.Methods The PBMC sample of 3 patients with sjgren syndrome and 3 healthy volunteers with consistent age were collected.The total RNAs was extracted from the PBMC samples,and reverse transcripted in vitro transcription(IVT),labeled with Cy5/Cy3 and hybridized on the gene chips.After scanning and data extraction with LuxScan 3.0,differentially expressed genes were analyzed with SAM method.The online tool of molecule analysis system(MAS) was used for biological knowledge mining.Results Statistical difference was calculated between the patient and control group in the following three pathways: cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway.Among these,genes of IL-2RA,IL-10 were up-regulated and genes of PF4,GZMA were down-regulated.Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjgren′s syndrome.And the research may provide new target therapy for SS.

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Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.

Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.