Objective To analyze CD+4 CDHi25 CDLo127 regulatory T cells (Treg) in peripheral blood of patients with thyroid nodules and differentiated thyroid cancer and their change regularity, and to investigate the immunosuppression mechanism. Methods The peripheral blood was collected from 175 patients with thyroid nodules, including 43 patients with differentiated thyroid cancer and 132 patients with nodular goiter.By using monoclonal antibodies, the blood samples were evaluated with the flow cytomertry for lymphocyte subsets and Treg cells. Results The results showed the prevalence of the CD+4 CDHi25 CDLo127 Treg in differentiated thyroid cancer group [(6.48±1.49) %] and nodular goiter group [(6.23+1.67) %] was higher than those in the healthy group [(5.62±1.48) %], and the difference was significant(P < 0.005), but there was no significant difference between the nodular goiter group and differentiated thyroid cancer group (P >0.005).Conclusion It is concluded that the relative increase of CD+4 CDHi25 CDLo127 Treg in peripheral blood of patients with nodular goiter and differentiated thyroid cancer may be related to immunosuppression and tumor progression.

OBJECTIVEThe present study was undertaken to design antisense oligodeoxynucleotides (AS-ODNs) of glial glutamate transporter-la (GLT-1a) and to evaluate the effectiveness of the designed AS-ODNs on the expression of GLT-1a.

METHODSFive sequences of GLT-1a AS-ODNs were designed according to the C terminus specific sequences of GLT-1a mRNA using antisense design software of IDT Com- pany. Western blot analysis was used to evaluate the inhibition effects of the five GLT-1a AS-ODNs on the expression of GLT-la.

RESULTSThe sequence of GLT-1a AS-ODNs with sequence of 5'-GGTTCTTCCTCAACACTGCA-3' could specifically inhibit the expression of GLT-1a in the hippocampal CA1 subfield of rats, while it had no effect on the expression of GLT-1b. This sequence showed similar inhibition on the expression of GLT-la in sham and ceftriaxone (Cef)-treated rats. It could also significantly inhibit the cerebral ischemic preconditioning (CIP)-induced up-regulation in the expression of GLT-1a. The magnitude of the inhibition in sham, Cef- or CIP-treated rats was similar by more than 60%.

CONCLUSIONFrom the designed five sequences of GLT-1a AS-ODNs, we obtained an effective sequence which can specifically inhibit the expression of GLT-1a.

Animals ; CA1 Region, Hippocampal ; metabolism ; Excitatory Amino Acid Transporter 2 ; antagonists & inhibitors ; metabolism ; Ischemic Preconditioning ; Oligonucleotides, Antisense ; genetics ; RNA, Messenger ; Rats ; Up-Regulation

Background Retinal microglia (RMG) plays an important role in the pathogenesis of retinal degenerative diseases,while chemokine CX3CL1 participates in the regulation of steady-state of microglia.It has been determined that bone marrow-derived mesenchymal stem cells (BMSCs) have a remarkable role to modulate the immune response and protect the central nervous system through the release of soluble factors in a paracrine fashion and further affect the functional behavior of cells.However,whether BMSCs are able to interact with RMG and activate related signaling pathway for the maintaining of homeostasis in the retina is still unclear.Objective The aim of this study was to investigate the interaction between BMSCs and lipopolysaccharide (LPS)-activated RMG in vitro,and dissect the effects of CX3CL1/CX3CR1 signaling pathway on the biological behavior of BMSCs and RMG.Methods RMG was isolated from SD rats,cultured with mixed culture of retinal glial cells and purified by shaking.The cells were identified by detecting the expression of CD111b,Iba1 and glutamamine synthetase (GS) with indirect immunofluorescence assay.LPS (1 mg/ml,2 μl) was added in the medium for 24 hours to stimulate RMG,and then the cells were divided into LPS control group,BMSCs group (cocultured with BMSCs for 24 hours) and CB-BMSCs group (cocultured with CX3CL1-blocking-BMSCs for 24 hours).The cells without LPS stimulation served as the blank control group.The functions of RMG,including the release content of tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β),the proliferation,phagocytosis,and migration of RMG were examined.Results RMG was successfully isolated and harvested from SD rats by using mixed culture of retinal glial cells and purified by shaking.CD11b and Iba1 showed the positive expression with the green fluorescence in the cells and GS was absent.The contents of TNF-αt in the cell supernatant were (2.55 ±0.97) ng/ml,(24.91 ±3.07) ng/ml,(20.38 ±2.97) ng/ml and (24.90 ± 1.88) ng/ml in the blank control group,LPS control group,BMSCs group and CB-BMSCs group,respectively,showing a significant difference among the groups (F=119.90,P＜0.05).The contents of IL-1 β in the cell supernatant were (1.12±0.36) ng/ml,(10.40±2.76) ng/ml,(7.00± 1.75) ng/ml and (9.55 ± 1.11) ng/ml in the blank control group,LPS control group,BMSCs group and CB-BMSCs group,respectively,showing a significant difference among the groups(F =34.96,P＜0.05).The secretory volume of TNF-α and IL-1 β were evidently lower in the BMSCs group than those in the LPS control group (both at P＜0.05),and no significant differences were found in the secretory volume of TNF-α and IL-1β between CB-BMSCs group and LPS control group (both at P＞0.05).The proliferative rate of RMG was lower in the BMSCs group than that in the LPS control group (P＜0.05),while there was no statistical difference between BMSCs group and CB-BMSCs group (P＞0.05).The mean fluorescence intensity (MFI) and the number of migrated RMG were considerably different among the four groups (F=70.55,15.49,both at P＜0.05),and those in the BMSCs group were significantly increased in comparison with the LPS control group (both at P＜0.05),while there was no significant difference between CB-BMSCs group and LPS control group (both at P＞0.05).Conclusions BMSCs could suppress the proliferation of LPS-activated RMG.Moreover,BMSCs might inhibit proinflammatory cytokines releasing,enhance phagocytosis and migration capabilities of RMG via CX3CL1/CX3CR1 signaling pathway.

Objective To measure the angular velocity and perpendicular ground reaction force of the ankle joint under different heights with half-squat jumping in parachute training simulation,providing a reliable experiment basis for the preventing of ankle injury.Methods A total of 18 volunteers participated in this study.The experimental group included 9 male with experience of parachute landing,while the other 9 male without experience of parachute landing were assigned to the control group.Each subject was instructed to jump off a platform with a height of 30 cm and 60 cm and land on a hard surface in a half-squat posture.The dynamic landing process was recorded with a high speed camera and the biomechanical data was collected and analyzed,including perpendicular ground reaction force,angular displacement,velocity and acting time.Results From 30 cm's height,the ankle angular displacement of the control group was significantly larger than the experimental group (25.73°± 8.13° vs 20.05°± 12.27°,P ＜ 0.05).The perpendicular ground reaction force of the control group was significantly smaller than the experimental group (3 372.4±748.6 N vs 5 181.5±1 726.2 N,P ＜ 0.05).The acting time of the control group was significantly longer than the ex perimental group (0.049±0.015 s vs 0.012±0.004 s,P ＜ 0.05).The buffer time of the control group was significantly shorter than the experimental group (1.397±0.746 s vs 1.737±0.451 s,P ＜ 0.05).From 60 cm's height,the ankle angular velocity of the control group was significantly higher than the experimental group (25.45± 15.01 °/s vs 16.51 ±4.18 °/s,P ＜ 0.05).The perpendicular ground reaction force of the control group was significantly smaller than the experimental group (4 616.0±1 124.7 N vs 7 119.5±2 307.4 N,P ＜ 0.05).The acting time of the control group was significantly longer than the experimental group (0.048±0.013 s vs 0.015±0.006 s,P ＜ 0.05).The buffer time of the control group was significantly shorter than the experimental group (0.922±0.347 s vs 1.617±0.547 s,P ＜ 0.05).Conclusion Jumping from different heights,the experinental group was larger in perpendicular ground reaction force but smaller in the angular velocity and displacement than the control group.There was a shorter acting time and a longer buffer time in the experimental group than the control group.

Objective To determine whether p38 mitogen-activated protein kinase (p38MAPK) signaling pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.Methods Two hundred and twenty-five male Kunming mice weighing 30-40 g were randomly divided into 4 groups:group control ( group C,n =55 ) ;group fractalkine (group F,n =60); group anti-CX3CR1 + fractalkine (group CF,n =55) and group SB203580 (p38MAPK inhibitor) + fractalkine (group SF,n =55).Fractalkine 100 ng was injected into cerebral lateral ventricle (i.c.v.) in groups F,CF and SF.Anti-CX3CR1 1 μg and SB203580 1 μg were injected i.c.v.at 1 h before fractalkine injection in groups CF and SF respectively.Paw withdrawal latency to a thermal nociceptive stimulus (PWL) was measured at 30 min before the drugs were injected into cerebral lateral ventricle and 30,60,120 and 240 min after fractalkine injection.Five animals were sacrificed after PWL measurement at each time point and their brains were removed for determination of phosphorylated p38MAPK protein expression (by Western blot analysis).Five animals were sacrificed at 30 min before the drugs were injected into cerebral lateral ventricle and 6,12 and 24 h after fractalkine injection for determination of IL-1β and TNF-α contents in the brain (by ELISA) in all the 4 groups.In group F 5 animals were sacrificed at 4 h after fractalkine injection for determination of action of fractalkine on microglia or astrocyte (by immunofluorescence).Results Fractalkine i.c.v.injection significantly reduced PWL and increased phosphorylated 38MAPK,IL-1β and TNF-α levels in group F as compared with group C.Pretreatment with anti-CX3CR1 or SB203580 significantly decreased fractalkine-induced hyperalgesia and phosphorylated-p38MAPK,IL-1β and TNF-α levels in groups CF and SF as compared with group F.Fractalkine was localized at microglia.Conclusion p38MAPK signal transduction pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.

OBJECTIVETo investigate the inhibitory effect of bone marrow mesenchymal stem cells (BMSCs) on E coliinduced prostatitis in rats.

METHODSBMSCs were isolated, cultured and amplified by the attached choice method. Fifty SD rats were randomized into five groups of equal number: normal control, acute bacterial prostatitis (ABP) , chronic bacterial prostatitis (CBP), ABP + BMSCs, and CBP + BMSCs, and the animals in the latter four groups were injected with E. coli into both sides of the prostate under ultrasound guidance for 1 - 14 days to induce ABP and for 4 - 12 weeks to induce CBP. The control rats were injected with the same amount of PBS. Two weeks after injection of BMSCs into the prostates, pathomorphological changes in the prostate were observed under the light microscope and the mRNA and protein levels of IL-1β and TNF-α determined by RT-PCR and ELISA, respectively, followed by statistical analysis with SPSS 18.0.

RESULTSHistopathological evaluation showed typical pathological inflammatory changes in the prostates of the rats in the ABP and CBP groups, including glandular structural changes, interstitial edema, inflammatory cell infiltration, and fibrous hyperplasia, which were all remarkably relieved after treated with BMSCs. The mRNA and protein levels of IL-β ([0.829 ± 0.121] and [271.75 ± 90.59] pg/ml) and TNF-α ([0.913 ± 0. 094] and [105.78 ± 19. 05] pg/ml) in the ABP and those of IL-1β ([0. 975 ± 0. 114] and [265. 31 ± 71. 34] pg/ml) and TNF-α ([0. 886 ± 0. 084] and [107. 45 ± 26. 11 ] pg/ml) in the CBP groups were significantly higher than those in the control rats ([0. 342 ± 0.087] and [45.76 17. 99] pg/ml, P <0. 05); ([0.247 ± 0.054] and ([19.42 ± 7. 75] pg/ml, P <0. 01) as well as than those in the ABP + BMSCs ([0. 433 ± 0. 072] and [51. 34 ± 22. 13] pg/ml, P < 0. 05 ) ; ( [0. 313 ± 0. 076] and [28. 38 ± 8. 78] pg/ml, P < 0. 01) and the CBP + BMSCs group ([0.396 ± 0.064] and [56.37 ± 21.22] pg/ml, P <0.05); ([0.417 ± 0.068] and [29.21 ± 10.22] pg/ml, P <0.01).

CONCLUSIONInjection of BMSCs can reduce E coli-induced prostatic inflammation reaction, which.may be associated with its reduction of inflammatory cell infiltration and the expressions of IL-1β and TNF-α in the prostate tissue.

Acute Disease ; Animals ; Bone Marrow Cells ; physiology ; Chronic Disease ; Escherichia coli Infections ; therapy ; Humans ; Interleukin-1beta ; genetics ; Male ; Mesenchymal Stromal Cells ; physiology ; Prostate ; metabolism ; Prostatitis ; metabolism ; microbiology ; therapy ; RNA, Messenger ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

The focus of microbiologists has moved to the rare actinomycetes.For selective isolation of rare actinomycetes that all play the important role in bioactive compounds,the approaches which involve the methods using gellan gum and flooding solution、 rehydration-centrifugation(RC)、 extremely high frequency radiation(EHF)、 bacteriophage and sucrose-gradient centrifugation were introduced in this paper.

The complete genomes of more cyanobacterial strains have been completed for sequence, and genetic engineering of cyanobacteria has evolved in the post-genome era. Since Kaneko and colleagues had completed the sequence for the complete genome of Anabaena sp. PCC 7120 in 2001, functions of some genes in this genome, including fructose-1,6-bisphosphate aldolase (FBA) gene, have been predicated using the method of bioinformatics. However, little information is available regarding whether this gene can encode FBA and its product characteristics of related enzyme. Here, to explore this information, the predicted II-FBA gene-encoding region in Cyanobase database was cloned by PCR method and then ligated into pET-32a to generate the expression vector, pET-FBA-II. The results of SDS-PAGE indicated that the expression level of the expected target polypeptide was approximate 23.4 percent as compared to total protein and the molecular weight is about 40 kDa as compared to the protein molecular marker. The results of enzyme activity analysis showed that the activity of II-FBA was ~11.8 U per mg protein and owned a standard activity of II-FBA. To sum up, the results not only prove the functional prediction of this II-FBA gene from the Cyanobase database, but also provide the important conditions for further studying its physiological and biochemical characteristics and functions of the gene expression product.

The synchronous electromyography of the extrinsic laryngeal muscles (sternohyoid muscle, sternothyroid muscle and thyrohyoid muscle) and thyroarytenoid muscle of dogs was analyzed in this study. The result showed that the electrical activity of the thyrohyoid muscle were synchronous to that of the thyroaryienoid muscle during quiet respiration, deep inspiration, phonation and swallowing. The discharge of the sternohyoid muscle was recorded during expiration and phonation. The electrical activity of the sternothyroid muscle was also recorded during inspiration and phonation, and was increased during deep inspiration. The study suggests that reinnervation of the thyroarytenoid muscle from thyrohyoid branch of the ansa cervicalis for functional rehabilitation could fit well the physiological characteristic of the larynx.

Objective To determine the effects and dosage of N?acetylcysteine( NAC) in the im?provement of the visibility in esophagogastroduodenoscopy( EGD) . Methods A prospective randomized con?trolled study was performed on 193 patients scheduled for EGD from November 2014 to July 2015 were ran?domized into five groups using digital table. In group A, 100 mg dimethicone and 2 g NaHCO3 were given. In group B,100 mg dimethicone, 2 g NaHCO3 and 20 000 U pronase were given. Group C received 100 mg dimethicone, 2 g NaHCO3 and 200 mg NAC. Group D received 100 mg dimethicone, 2 g NaHCO3 and 400 mg NAC and group E 100 mg dimethicone, 2 g NaHCO3 and 600 mg NAC.The agents were dissolved in 100 ml water for each patient.Endoscopy was completed by one endoscopist and the score of image visibility assessment was completed by 2 other endoscopists. The 3 endoscopists were unaware of grouping. The total scores, the time of washing, the time of examination and complications were compared and analysed. The total image scores of group A, B, C,D and E were 30?83±3?78, 35?69±2?88, 33?16±3?90, 34?95±3?46 and 36?76±2?91, respectively. Group A was the lowest(P<0?05),followed by group C(P<0?05). There were no differences among group B,D, and E(P>0?05).Images that were scored 3 were the most in group E.The washing times of each group were 38?00±19?10, 17?03±11?44, 15?92±10?81, 15?78 ±10?24 and 15?55±9?69, and the examination times of each group were 13?49±2?49, 9?41±1?86, 9?08± 1?80, 8?73±1?91 and 8?78±1?79 minutes.Group A was the longest in these two indices(P<0?05). There were no significant differences among group B, C, D and E ( P<0?05) . There were no significant differences in adverse effects among groups after EGD( P>0?05) . Conclusion The preoperative NAC can improve the visibility in EGD. The best dose is 600 mg, whose effects and safety were similar to those of 20 000 U, but yield to less washing time and the examination time in EGD.