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.

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 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.

ObjectiveTo investigate the value of combined digital mammography and Color Doppler Mammasonography in the diagnosis of the breast carcinoma with ROC analysis.MethodsFifty female patients with breast carcinoma and fifty female patients with breast benign lesions confirmed by means of surgery and histopathological examinations were studied.All patients were examined by Digital Mammagraphy and Color Doppler Mammasonography.Images of all patients were sorted and divided into three groups,Digital radiographs of all patients was the first group,and the second group was Color Doppler photographs,and the third was included the above two photographs.Each group was evaluated respectively by three radiologists independently.The diagnostic result of breast carcinoma or benign lesions were investigated with a five point confidence-rating scale:(1)definitely positive;(2)probably positive;(3)indeterminate;(4)probably negative,and(5)definitely negative.ROC analysis was made with SPSS 11.0 software.The differences of the area under the curve for each group was made by Z test.ResultsThe average number of breast carcinoma diagnosed by three radiologists with digital mammography,Color Doppler Mammasonogaraphy and both of them was 42,38,and 48,respectively;and the average number of breast benign lesions was 40,43,and 47,respectively.The sensitivity and specificity for digital mammography was 84%(42/50),80%(40/50),and they were 76%(38/50),86%(43/50)for Color Doppler;and 96%(48/50),94%(47/50)for two modes combination.ROC analysis indicated that the average area under the curve was 0.852 for Digital Mammagraphy,0.825 for Color Doppler,and 0.952 for both combined.For digital mammography combined with Color Doppler Mammasono-garaphy on diagnosis breast carcinoma,the area under the curve(Az)was significally larger than that for digital mammography(Z1=2.183,P＜0.01),and also larger than that for Color Doppler Mammasono-garaphy(Z2=2.581,P＜0.01), respectively.Conclusions Among three medical imaging modes on diagnosing breast carcinoma,the diagnostic value of digital mammography combined with Color Doppler Mammasono-garaphy was the highest. It was helpful on detecting and diagnosing breast carcinoma early.

Objective To investigate etiology of acute pneumonia in children in order to provid basis for clinical diagnosis and treatment.Methods The children with acute pneumonia who were hospitalized in children's hospital affiliated to Suzhou university were selected.And the sputum of them were collected.Bacteria and virus were tested using sputum culture and direct immunofluoresence respectively.Antibodies against mycoplasma and chlamydia were detected by enzymelinked immunosorbent assay(ELISA) in paired sera. Results Microbial etiology was obtained in 360 cases (67.7%) of 532 patients.Viral infections were in 178 cases (33.5%).Bacterial infections were in 23 cases (4.3%),mycoplasma pneumoniae 50 cases (9.4%) and chlamydia pneumoniae 19 cases (3.6%),compound infections 90 cases (16.9%).Respiratory syncycial virus was the major viral pathogen,streptococcus pneumoniae were the prominent pathagens of bacterial pneumonia,followed by haemophilus influenza.Conclusions Viral infection is the most common cause of acute pneumonia in children in Suzhou area during winter and spring,followed by mycoplasma pneumoniae,bacteria and chlamydia pneumoniae.Mycoplasma pneumoniae infection is more common in children older than 3 years,chlamydia pneumoniae infection is more in infants less than 3 months,most of compound infection children were below the age of 3 years.

Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

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.