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.

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

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.

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.

AIM: To study the effect of Ginkgo biloba extract (GbE) on the expression of connective tissue growth factor (CTGF) in the initiating stage of pulmonary fibrosis of rats after administration of bleomycin (BLM).METHODS: The expression of CTGF in lungs was detected by Western blotting. The content of hydroxyproline was assayed by the method of chloramines T. The content of malondialdehyde (MDA) in plasma was investigated by colorimetry.RESULTS: On day 14 after administration of BLM, the contents of CTGF in lungs and MDA in plasma in BLM+NS group were higher than those in NS group, respectively (P

Objective To explore the application of 18F-FDG PET/CT combined with high-resolution CT (HRCT) in the same scanner on diagnosis of lung cancer, and its influencing factors. Methods 50 patients, in which some cannot supply HRCT raw date and under highly suspicion of being lung cancer, some were postoperative lung cancer and metastasis of lung cancer, were examined by 18F-FDG PET/CT combined with HRCT in the same scanner. Results 50 patients were all successful (100%) on PET/CT scans after preparation, injection, rest and breathing exercises;46 cases (92%) were successful on PET/CT combined with HRCT scans;4 cases (8%) failed on HRCT scans. Ma-lignant lesions were found in 35 cases, with the metastasis of 21 cases;4 in 6 cases of postoperative lung cancer were found metastasis;9 cases of benign pulmonary nodules need to be observed sequentially. Conclusion 18F-FDG PET/CT combined with HRCT in the same scan-ner is valuable on diagnosis of lung cancer. The diagnostic accuracy and sensitivity significantly increase. It is a non-invasive new imaging technology and systemic metastasis can be observed.

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

To retrospectively analyze the safety and efficacy of low dose subcutaneous decitabine regimen in patients with acute myeloid leukemia (AML) and intermediate-or higer-risk myelodysplastic syndrome (MDS).Of 6 AML cases,2 achieved complete remission (CR),2 with partial remission(PR),1 with stable disease(SD),1 with progressive disease(PD).As to the 8 MDS patients,one achieved CR and 6 with hematologic improvement (HI),1 case SD.Low dose subcutaneous decitabine regimen could be an alternative choice of older AML or MDS patients.

Objective: To observe the clinical efficacy of grain-sized moxibustion in treating chemotherapy-induced myelosuppression for non-small cell lung cancer (NSCLC) and its effect on quality of life (QOL). Methods: Eighty NSCLC patients admitted to the Inpatient Department of Zhejiang Cancer Hospital between September 2016 and March 2018 were recruited and divided into an observation group and a control group by random number method, with 40 cases in each group. The two groups both received chemotherapy with paclitaxel plus cisplatin (TP regimen). The control group received oral administration of leucogen tablets starting from the first day of chemotherapy, 20 mg each time, three times a day, for consecutive 14 d; the observation group was additionally given grain-sized moxibustion, once a day, five days per week at a two-day interval, until the fourteenth day. The myelosuppression severity was observed and compared between the two groups prior to chemotherapy, at the 3rd, 7th and 14th days of chemotherapy; the QOL in the two groups was evaluated before chemotherapy, at the 14th and 21st days of chemotherapy. Results: Regarding myelosuppression, the peripheral blood indicators increased significantly at the 3rd day of chemotherapy in both groups (P<0.05 or P<0.01); at the 7th and 14th days of chemotherapy, the peripheral blood indicators presented a decreasing tendency in the two groups, but the level in the observation group was still significantly higher than that before chemotherapy (P<0.01); at the 3rd, 7th and 14th days of chemotherapy, the peripheral blood indicators in the observation group were higher than those in the control group (P<0.05 or P<0.01); the occurrence rate of myelosuppression in the observation group was significantly lower than that in the control group (P<0.01). The QOL score in the observation group was markedly higher than that in the control group at the 14th and 21st days of chemotherapy (both P<0.05). Conclusion: Grain-sized moxibustion can effectively improve myelosuppression after chemotherapy for NSCLC, reducing its occurrence and enhancing the patient's QOL.