Objective To construct small hairpin RNA (shRNA) expression plasmid targeting rat opticin gene. Methods Four pairs of opticin oligonucleotides were synthesized and inserted into the plasmid vector, resulting into four plasmids: shRNA-1, shRNA-2, shRNA-3 and shRNA-4. Then the four constructed shRNA expression vectors and empty vector were transfected into rat ciliary non-pigment epithelium (NPE) cells by lipofectmaine 2000. Non-transfected NPE cells were set as control group. The expression of opticin mRNA and protein were measured by Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot respectively. Results The opticin mRNA expression of the shRNA-1,shRNA-2, shRNA-3, shRNA-4 group were decreased compared with the control group (F = 10. 239, P = 0. 000);the inhibitory rate were 85.7% ,62. 87% ,54.87% and 48.77% respectively. The opticin protein expression of the shRNA-1,shRNA-2, shRNA-3, shRNA-4 group were also decreased compared with the control group (F=17.870, P= 0.000);the inhibitory rate were 78.7%, 34.6%, 31.1% and 16.8% respectively.Conclusions The shRNA-1 expression plasmid has most potent inhibitory effect on opticin expression in rat ciliary NPE cells.

OBJECTIVE:To study the separation and purification of polysaccharides sulfate from Ulva lactuca,and to analyze its structure.METHODS:The separation and purification method of polysaccharides sulfate from U.lactuca was optimized.The content of polysaccharides in polysaccharides sulfate was determined by phenyl-sulfuric acid method.The molecular amount of polysaccharides was determined by HPGC.The structure of polysaccharides was analyzed by IR and 13C-NMR.RESULTS:The optimal extraction condition of polysaccharides sulfate from U.lactuca were crashed materials 20 mesh sieve,20-fold water,decocting 3 times at 85 ℃,2 hours each time.The content of polysaccharides was 22.64% and molecular amount was 1 156 112.Soluble polysaccharides mainly contained ?-glucuronic acid,rhamnose and plenty of sulfate radical.CONCLUSION:The method is simple and stable,which provides theoretic basis for production of polysaccharides sulfate from U.lactuca.

Objective: To establish a determination method for the enantiomer in safinamide mesilate. Methods: A Chiralpak ASH (250 mm ×4. 6 mm, 5 μm) column was used with the mobile phase of n-hexane-ethanol-diethylamine (75 ∶ 25 ∶ 0. 1). The flow rate was 1. 0 ml·min-1 . The wavelength was set at 225 nm. The column temperature was 35℃. Results: The resolution of safinamide mesilate and the enantiomer was above 2. 0. The linear range of them was 1. 007-2 517. 500 μg·ml-1 and 0. 909-2273. 200 μg·ml-1 ,respectively(r = 0. 999 0). The average recovery of the enantionmer was 104. 9% with RSD of 2. 3% (n = 9). Conclusion: The method is accurate and rapid, and suitable for the determination of the enantiomer in safinamide mesilate.

Objective:To establish a method for the determination of enantiomer in rivaroxaban by HPLC. Methods:A Chiralpak IA column (250 mm × 4. 6 mm, 5 μm) was used with the mobile phase of 100% methanol. The flow rate was 1. 0 ml·min-1 . The detection wavelength was set at 250 nm and the column temperature was 40℃. Results:The resolution of rivaroxaban and the enanti-omer was above 2. 0. The linear range of them was 0. 5-1 000. 00 μg·ml-1(r=0. 999 9). The detection limit of the enantiomer was 0. 17ng and the average recovery was 102. 9% with RSD of 1. 54%(n=6). Conclusion:The method is accurate and rapid, and suit-able for the determination of the enantiomer in rivaroxaban.

Objective To investigate the effects of hyperbaric oxygen (HBO) on the expression of caveolin2 and matrix-metalloproteinase-9 (MMP-9) in brain tissues and the blood brain barrier (BBB) after cerebral ischemia and reperfusion (I/R) in rats. Methods Three hundred and seventy male Wistar rats were randomly assigned into sham,ischemia-reperfusion,HBO,and I/R + HBO groups. After creating cerebral I/R models,oxygen at 0.25 MPa was administered 5 times,and 2％ Evans blue (EB) was injected into the tail veins 1 h before the rats were sacrificed.The permeability of the BBB,the expression of caveolin-2 and MMP-9,and EB content were determined by Western blotting,immunohistochemistry,and spectrophotometery,respectively. Results In the I/R group,the EB content increased steadily to a peak at 4 hours.EB content in the IR + HBO group was significantly lower than in the I/R group.Caveolin-2 and MMP-9 were significantly augmented by I/R injury at the 24th,48th and 72nd hours.Compared to the I/R group,HBO intervention decreased their expression levels. Conclusion HBO intervention can reverse the increase of caveolin-2 and MMP-9 caused by I/R injury,which suggests a mechanism for protective effects of HBO on the permeability of the BBB in I/R injury.

BACKGROUND:Mesenchymal stem cells are found to have the immunoregulatory activities and a potential application prospect in the treatment of autoimmune diseases.
OBJECTIVE:To explore the mechanism of transplanting mesenchymal stems cells on the treatment of multiple sclerosis.
METHODS:The mouse mesenchymal stems cells were prepared, and injected into the al ogenic and syngenic normal mice, to detect the frequency of CD4+CD25+Foxp3+T cells in the spleen, thymus, and lymph nodes by flow cytometry, and to detect the Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes by reverse transcription-PCR.
RESULTS AND CONCLUSION:Transplantation of mesenchymal stem cells on normal mice led to a significant up-regulation of CD4+CD25+Foxp3+T cells, Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes both in the al ogenic and syngenic transplant groups. Transplantation of mesenchymal stem cells may be an available method in the treatment of autoimmune diseases, and CD4+CD25+Foxp3+T cell, Foxp3, transforming growth factor-β1, and interleukin-10 may be involved in this process.

OBJECTIVETo compare the difference of clinical efficacy on sleeping disorder in the children with encephalopathy between the combined therapy of acupuncture at head points and seed-pressure at ear points and the simple acupuncture at head points.

METHODSThirty cases of sleeping disorder induced by encephalopathy werei randomized into an observation group and a control group, 15 cases in each one. In the observation group, the combined therapy of acupuncture at head points and seed-pressure at ear points was adopted. The head points in cluded Sishencong (EX-HN 1), Shenting (GV 24) and Benshen (GB 13). The ear points were the positive reactive sites in the cymba and cavum conchae. In the control group, acupuncture was applied simply to the acupoints on the head. The treatment was given once on every Tuesday and Friday a week separately, 30 min each time. Totally, 16 treatments were required. Children's sleeping habit questionnaire (CSHQ) was used to observe the sleep improvements and the efficacy in the patients of the two groups.

RESULTSIn the observation group, the results of sleep resistance, sleep anxiety, night sleep wake, parasomnias, sleep dyspnea, daytime somnolence and the total score after treatment were all improved apparently as compared with those before treatment (all P<0. 05). In the control group, the results of night sleep wake, parasomnias, daytime somnolence and the total score after treatment were improved apparently than those before treatment (all P<0. 05). In the observation group, the results of sleep resistance, sleep dyspnea and the total score after treatment were better than those in the control group (all P<0. 05) and the scores of sleep anxiety and daytime somnolence in the control group were better than those in the observation group after treatment (both P<0. 05).

CONCLUSIONThe combined therapy of acupuncture at head points and seed-pressure at the positive reactive sites in the cymba and cavum conchae achieves the superior efficacy on sleep resistance and sleep dyspnea as compared with the simple acupuncture. The efficacy of simple acupuncture is more satisfactory on sleep anxiety and daytime somnolence.

Acupuncture Points ; Acupuncture Therapy ; Acupuncture, Ear ; Brain Diseases ; complications ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Sleep ; Sleep Wake Disorders ; etiology ; physiopathology ; therapy

Objective To explore the effect of weightlessness simulation on processing speed of mental rotation of males in 72 h head-down tilt,in order to further reveal human performance changes caused by weightlessness.Methods During 72 h weightlessness simulation (9 am day 1 to 9 pm day 4,head-down tilt started from 9 pm day 1),16 male subjects were processed speed of rotation data byhand picture mental rotation (test 8 times),using self cross-references design,and the variation of processing speed of mental rotation by duration was analyzed.Results The linear regression analysis showed,the reaction time of each time point for each subject as the independent variable,the angle of rotation as the dependent variable,with the extension of simulated weightlessness time.The intercept(constant term of the regression equation),indicating the processing speed of non-rotation,were gradually increased with HDT time,but didn(t) reached significantly difference (F(3,14) =0.551,P =0.650) ;the slope(Regression coefficient of the regression equation),indicating the processing speed of rotation.were gradually increased with HDT time,and reached the significantly difference (F(3,14) =4.338,P =0.009).The value of intercept and slope was like upside-down U in three days,and reached the highest value at day 2.Conclusion Weightlessness simulation have affection on the speed of mental rotation,especially on the rotation speed.

Objective: To observe the clinical effect of acupuncture treatment for infantile autism. Methods: After being treated with the needling technique for enhancing wisdom and opening the orifice, 32 cases of sick children were assessed by Autism Treatment Evaluation Checklist (ATEC) depending upon their clinical symptoms. Results: After acupuncture treatments, language communication, interpersonal relationships, movements and behavior, and personal ability had all improved. Among them, the remarkable rate of language communication showed a statistical significance in comparison with the other three items (P<0.05). In the comparison of the ATEC scores before and after the treatments, the language items (P<0.01), social relationships, perception and behavior items, showed statistically significant differences (P<0.05). Conclusion: Acupuncture therapy can effectively improve the clinical symptoms of sick children with autism, and the interference effect on language function is especially remarkable.

Objective To investigate the pathological mechanism of delayed endolymphatic hydrops(DEH) , and clarify the clinical value of endolymphatic space imaging after intratympanic injection of gadolinium in the diag_nosis of delayed endolymphatic hydrops .Methods Twenty -four hours after bilateral intratympanic injection of gadolinium ,the locations and severity of endolymphatic hydrops of all patients were evaluated by using three dimen_sional fluid-attenuated inversion recovery (3D -FLAIR) and three dimensional real inversionrecovery (3D -real IR) .ResuIts All patients had unilateral or bilateral endolymphatic hydrops .Among 9 ipsilateral DEH patients , only 1 (11 .1% ) patient was identified as mild endolymphatic hydrops and the rest (88 .9% ) examined had signifi_cant endolymphatic hydrops in vestibule of their affected ears ;Endolymphatic hydrops appeared in cochlea of the af_fected ear in 8 (88 .9% ) patients ,except for 1 patient .Endolymphatic hydrops were not observed in the contralater_al ears of 9 ipsilateral DEH patients .Mild endolymphatic hydrops in bilateral vestibule ,severe in right cochlear and none in left cochlea of contralateral DEH patient were identified .ConcIusion Endolymphatic hydrops is the primary pathological factors of DEH .Endolymphatic space imaging after intratympanic injection of gadolinium can intuitively reflect the locations and severity of endolymphatic hydrops in DEH patients .