To counteract the immune system in parasitic hosts, some viruses encode proteins to suppress the RNA interference (RNAi) effect. In this report, we established two RNAi systems to be easily observed with strong and obvious effect. The function of the P19 of tomato bushy stunt virus, which suppresses RNAi in mammal cells, was then studied using these two systems. Short hairpin RNAs targeting green fluorescence protein (pshRNA-GFP) and firefly luciferase (pshRNA-luc) were designed and inserted into a eukaryotic transcriptional vector pTZU6+1, respectively. The shRNA expressing vectors were co-transfected with plasmids containing the target gene with or without P19. The GFP expression level was assayed by fluorescence microscopy, Western blotting and RT-PCR. The luciferase expression level was analyzed by the dual-luciferase assay system. pshRNA designed in this study down-regulated the target gene specifically and efficiently, with a decrease of expression of both genes of about 70%, respectively. When P19 was introduced into the RNAi systems, the expression of both GFP and the luciferase were mostly recovered compared with the control groups. The RNAi systems of GFP and luciferase were constructed successfully, demonstrating that P19 of tomato bushy stunt virus has the ability to counteract the RNAi effect induced by shRNA in mammal cells.

AlM: To monitor long - term changes of matrix metalloproteinase-2 ( MMP-2 ) in human tears fluid after laser in situ keratomileusis ( LASlK) .
METHODS: Thirty - two myopia cases ( 64 eyes ) underwent uneventful LASlK were enrolled in the study. Tear fluid were collected and MMP-2 expression was analyzed by Western - bolt assay preoperatively and postoperatively on 15d, at 1, 3mo, and 1a.
RESULTS:LASlK increased the concentration of MMP-2 in human tear fluid. At 15d postoperatively, the magnitude of MMP-2 was 1. 4 times that of preoperative, thereafter subsided, but didn't return to preoperative level by 3mo ( P < 0. 05 ). Up to 1a after surgery, the concentration of MMP-2 almost recovered (P>0. 05).
CONCLUSlON: MMP- 2 is significantly expressed in human tear fluid after LASlK, then subsided with time, but didn't return to preoperative level by 3mo and almost recovered up to 1a, indicating wound healing of LASlK would continue up at least 3mo after surgery and almost recovered 1a postoperatively.

Adenoma, Villous ; metabolism ; pathology ; surgery ; Aged ; Diagnosis, Differential ; Female ; Humans ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Mullerian Ducts ; pathology ; Papilloma ; pathology ; Vaginal Neoplasms ; metabolism ; pathology ; surgery

Objective To evaluate the effect of hydrogen-rich saline on inflammatory responses during transient cerebral ischemia-reperfusion (I/R) in rats.Methods Forty-five male Sprague-Dawley rats,aged 5-6 yr,weighing 180-250 g,were randomly divided into 3 groups (n =15 each) using a random number table:sham operation group (group S),group I/R and hydrogen-rich saline group (group H).The rats were anesthetized with intraperitoneal 10％ chloral hydrate 0.3 ml/100 g.Transient cerebral ischemia was induced by bilateral common carotid artery occlusion with hypotension for 15 min,followed by reperfusion.Five rats were randomly chosen from each group,and Morris water maze was used to assess the cognitive function starting from 5 days before establishment of the model.Place navigation test lasted for 5 consecutive days.The escape latency,swimming speed and swimming distance were recorded.Spatial probe test was carried out on 1 and 3 days after establishment of the model.The time of staying at the target platform quadrant,frequency of crossing the original platform and swimming speed were recorded.The rats were sacrificed after the end of spatial probe test on 3 days after the model was established,and hippocampi were removed to examine the morphology of neurons in hippocampal CA1 region with light microscope.Five rats randomly chosen from each group were sacrificed on 1 day after the model was established,and hippocampi were removed to detect the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β).The rest 5 rats in each group were sacrificed,and hippocampi were removed for determination of nuclear factor kappa B (NF-κB) activity (by immuno-histochemistry).Results In place navigation test before the model was established,the escape latency and swimming distance were gradually shortened with the prolonging training time,and no significant change was found in the swimming speed with the prolonging training time in the three groups.Compared with group S,the time of staying at the target platform quadrant was significantly shortened,and the frequency of crossing the original platform was reduced on 1 and 3 days after establishment of the model,and the contents of TNF-α and IL-1β and NF-κB activity were increased on 1 day after establishment of the model in group I/R.Compared with group I/R,the time of staying at the target platform quadrant was significantly prolonged,and the frequency of crossing the original platform was increased on 1 and 3 days after establishment of the model,and the contents of TNF-α and IL-1β and NF-κB activity were decreased on 1 day after establishment of the model in group H.There was no significant change in the swimming speed during spatial probe test on 1 and 3 days after establishment of the model.Conclusion The mechanism by which hydrogen-rich saline reduces transient cerebral I/R injury may be related to inhibition of inflammatory responses in rats.

To investigate formation mechanism of secologanic acid sulfonates in sulfur-fumigated buds of Lonicera japonica, secologanic acid was enriched and purified from the sun-dried buds of L. japonica by various column chromatography on macroporus resin HPD-100, silica gel and ODS. The stimulation experiments of sulfur-fumigation process were carried out using secologanic acid reacted with SO2 in the aqueous solution. The reaction mechanism could be involved in the esterification or addition reaction. The present investigation provides substantial evidences for interpreting formation pathway of secologanic acid sulfonates in sulfur-fumigated buds of L. japonica.
Alkanesulfonates ; chemistry ; Carboxylic Acids ; chemistry ; Chromatography, High Pressure Liquid ; Flowers ; chemistry ; drug effects ; Lonicera ; chemistry ; drug effects ; Models, Chemical ; Molecular Structure ; Sulfur ; chemistry ; pharmacology ; Sulfur Dioxide ; chemistry ; Water ; chemistry

Prolactin is a polypeptide hormone which mainly acts on the reproductive system and plays an important role in penile erection and ejaculation. Prolactin receptors have a variety of short forms apart from the classic long form, which are widely expressed in male reproductive glands. High levels of prolactin can induce erectile dysfunction and results in secondary male infertility, which are mainly associated with the inhibition of dopaminergic activity, reduction of the testosterone level, and contraction of the cavernous smooth muscle. Moreover, low levels of prolactin can result in ejaculatory dysfunction. This article updates the views on the expressions of prolactin receptors in the male reproductive system, the effects of prolactin on penile erection and ejaculation, and its action mechanisms.
Ejaculation ; physiology ; Erectile Dysfunction ; Humans ; Infertility, Male ; Male ; Muscle, Smooth ; physiopathology ; Penile Erection ; physiology ; Prolactin ; physiology ; Receptors, Prolactin ; physiology ; Reproduction

Objective Toinvestigate the metabolic characteristics of triglyceride (TG) in the plasma of rabbitswith steroid?induced osteonecrosis of the femoral head and the therapeutic effect of Gushiling Capsule. Methods Rabbit model of femoral head necrosis induced by hormone was established. The level of plasma TG was detected by HPLC/MS and the relationship between the changes of plasma TG level and local histomorphology of femoral head was analyzed. Results Compared with the normal control group, significant changes of plasma TG levels were observed in the model group, the traditional Chinese medicine group and the western medicine group at each time point (P<0.01). However, no significant difference was found between the traditional Chinese medicine group and the western medicine group. Compared with the model group, only a small amount of high signal joint imaging and central signal of the femoral head were found by the MRI examination in the traditional Chinese medicine group and the western medicine group. Pathology assay results showed the degree of femoral head osteonecrosis in the Chinese medicine group was milder than that in the Western medicine group. Conclusion Gushiling Capsule and western medicine Simvastatin have the similar effect of reducing plasma TG, which may be one of the mechanisms underlying Gushiling Capsule preventing steroid?induced femoral head necrosis.

Objective To observe the clinical response and safety of second-generation tyrosine kinase inhibitor dasatinib in the treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph + ALL). Methods The clinical data of 10 adult Ph + ALL patients treated with dasatinib were analyzed with review of literatures. Results All the 10 Ph+ ALL patients treated with dasatinib achieved remission in 7 weeks, including 9 cases of complete remission [7 cases achieved complete molecular remission (CRm) in 13 weeks]. The median overall survival time (OS) was 13.8 months (5-33), and the median disease-free survival (DFS) time was 10.8 months (4-25). There were 3 cases of pleural effusion, 4 cases of Ⅳ degree of bone marrow suppression and 6 cases of extremely low blood platelet, which could be that was improved via by symptomatic treatment and, with no case of the death occurred during the treatment of dasatinib, the safety was high. Conclusion Dasatinib can deepen molecular biological reaction and prolonged the survival time of patients in the treatment of adult Ph+ ALL, with high remission rate and safety, and which can be considered as first-line treatment.

Objective To observe the effectiveness and side-effect of two cases of relapsed and refractory T-cell lymphoma (TCL) treated with thalidomide and interferon.Methods Two cases of relapsed and refractory TCL was treated with thalidomide and interferon, the efficacy and side-effect were observed, and the relevant literature was reviewed.Results The patients achieved partially remission after being treated with thalidomide and interferon.Conclusion Thalidomide in combination with interferon can be used as a second line therapy for relapsed and refractory TCL.

Aim To investigate the effect of 2-deoxy-D-glucose(2-DG)on the sensitivity of leukemia multi-drug resistant K562/ADMcells to adriamycin by inhib-iting glycolytic pathway as well as its molecular mecha-nisms.Methods The leukemia drug-resistant K562/ADM cells and parental K562 cells were used as the target cell models.The cell proliferating activity was assessed with an MTT colorimetric assay,and the gly-colysis including glucose consumption,lactate export, and hexokinase activity was determined by glucose, lactic acid and hexokinase (HK)testing kits.The ex-pression and phosphorylation of mammalian target of rapamycin(mTOR)and glucose transporter-4 (GLUT-4)expression were analyzed by western blot.Results K562/ADM drug-resistant cells possessed higher HK activity,GLUT-4 expression level and aerobic glycolic ability than K562 sensitive cells. 2-DG treatment markedly inhibited HK activity,glucose consumption, and lactate export both in K562 cells and K562/ADM cells,and suppressed the proliferation of the two cells in a time-and concentration-dependent manner.Low concentration of 2-DG or adriamycin could increase the expression and phosphorylation of mTOR.However, the co-administration of 2-DG and adriamycin markedly counteracted adriamycin-mediated enhancement of mTOR expression and phosphorylation and down-regu-lated GLUT-4 expression in K562/ADM cells,and 2-DG dramatically improved the sensitivity of K562/ADM cells to cytotoxicity.Conclusion 2-DG inhibits the proliferation of drug-resistant K562/ADM cells and en-hances the sensitivity to adriamycin by blocking aerobic glycolysis pathway through inhibiting hexokinase activi-ty,counteracting adriamycin-stimulated increased ex-pression and phosphorylation of mTOR and downregu-lating GLUT-4 expression.