Objective To investigate the inhibition of pathogenic viral gene expression in HPV6bE6-positive cell line by specific siRNA, which might have great potential for clinical use. Methods B16 cells were transfected with recombinant plasmid pcDNA3.1(+)-GFP/HPV6bE6, and the positive cell clones were selected by fluorescence protein observation and RT-PCR. Four specific siRNAs, none of which shares homology with exons of known human genes, were designed and synthesized to target HPV6bE6 mRNA. Quantitative real-time PCR was performed to measure the inhibition rates of target gene expression by comparing HPV6bE6 mRNA concentrations before siRNA transfection with those after transfection. The inhibition rates of target gene expression with different siRNA concentrations of 0.2 nmol/L, 1 nmol/L, 10 nmol/L, 50 nmol/L, 150 nmol/L and with different treatment time at 24 h, 48 h, 72 h and 96 h after transfection were measured respectively. Results More than 90% reduction of HPV6bE6 mRNA was observed following treatment with HPV6bE6-siRNA, and HPV-negative cells were apparently unaffected by HPV6bE6-siRNA. The decrease of HPV6bE6 mRNA was maximal at 24 h after siRNA treatment and sustained for at least 4 days. The minimal level of siRNA to efficiently silence the homogeneous target gene HPV6bE6 was 1 nmol/L. Conclusion HPV6bE6-siRNA can efficiently and specifically silence target genes and may be developed as a potential therapeutic approach for HPV infection.

Objective To investigate the incidence of nutritional risk for stroke patients with diabetes mellitus. Methods All the stroke patients with diabetes mellitus in a neurologic department were investigated with Nutritional Risk Screening (NRS2002). Results The incidences of undernutrition and nutritional risk were 11.3% (42/372) and 35.8% (133/372), respectively. The incidence of nutritional risk was more in the patients aged over 60 years than in the patients below 60 years (P=0.001). The incidence of both undernutrition and nutritional risk was more in the patients with stroke relapsing than those of the initial stroke (P<0.001). Conclusion The stroke patients with diabetes mellitus are in the risk of undernutrition and nutritional risk, especially those over the age of 60 years and relapsed stroke.

Objective To investigate the efficacy and safety of autologous cryopreserved platelet transfusion in the management of thrombocytopenia after chemotherapy in hematological malignancy.Methods A total of 40 patients diagnosed as hematological malignancy with complete remission were equally assigned into study group and control group.During chemotherapy interval in the study group,when platelet counts exceeded 120 × 109/L,autologous platelets were collected with CS3000 Cell Separator and cryopreserved at-80℃ with 5％ dimethylsulfoxide.When platelet counts dropped below 15 × 109/L after chemotherapy,autologous platelets were thawed with 40℃ water bath and transfused back to each patient.In the control group,when platelet counts dropped below 15 × 109/L after chemotherapy,allogeneic fresh platelets were transfused.Median loss during the freeze-thaw-wash procedure in study group was observed,and the 1 h,24 h corrected count increments(CCI)were calculated in the both groups.The hemostatic effects and adverse reactions were also observed.Results In the control group,1hCCI and 24hCCI were (19.3 ±6.1)× 109/L and(12.2 ± 7.0)× 109/L,respectively,with the effective rate of 80％ and the transfusion reaction rate of 45％.Totally 20 collection and transfusions were finished in the study group.A total of(3.4-8.5)× 1011 platelet were obtained in each collection.Platelet recovery after freezing and thawing was(73.51 ±9.03)％(62％-83％).1hCCI was(17.4±7.6)× 109/L,24h CCI was(10.5 ±5.8)× 109/L and the effective rate was 85％.There was no significant different between the two groups (P ＞ 0.05).The transfusion reaction rate was 15 ％,which was significantly lower than that of the control group(P ＜ 0.05).Meanwhile,adverse reactions were occurred less in the study group.Conclusion This study demonstrates that autologous cryopreserved platelet transfusions can be safely administered for supporting thrombocytopenia in hematological malignancy patients undergoing chemotherapy.

Objective To establish a radioresistant cell subline from a human A549 lung cancer cell line and investigate the mechanism of radioresistance. Methods Two proposals were applied for the non-small cell lung cancer A549 cells irradiated with X-rays:A group of A549 cell line was irradiated five times, the fractionated dose was 600 cGy, and the other group was exposed 15 times, the fractionated dose was 200 cGy. After the completion of irradiation, two monoclones were obtained from the survival of cells and named the subline A549-S1 and A549-S2. The radiosensitivity and cell cycle distribution of these two clones,together with its parental A549 cells were measured by clone formation assay and flow cytometry.The mRNA and protein levels of Notch1 in A549 cell line and the sublines were determined by RT-PCR and Western-blots. Results Compared with the parental A549 cells, A549-S1 cells showed significant resistance to radiation with D0, Dq and N values increased, and a broader initial shoulder as well as 1.38-fold increased value of SF2. The A549-S1 subline also showed higher percentage of cells in S phase and G2/M phase, but lower percentages in G0/G1 phase (P<0.05). The expression of Notch1 in A549-S1 was enhanced obviously than in A549 cells. But for A549-S2 the radioseasitivity was slightly increased compared with the parental cells with D0, Dq and N values decreased and a curve initial shoulder. The ratio of cells in S and G0/G1 phase ratio was lower than that in parental A549 cells, but that in G2/M phase ratio was higher significantly (P<0.05) .The expression of Notch1 had no marked change compared to A549 cell. Conclusions The radioresistance of the A549 cell subline is correlated with the irradiation program. The cell subline shows a different cell cycle distribution from their parental line. The cell cycle distribution has a close correlation with the expression of Notch1.

AIMTo study the chemical constituents of Salsola collina Pall..

METHODSCompounds were isolated by silica gel column chromatography. IR, MS, 1HNMR, 13CNMR, HMQC, HMBC, DEPT were used for the structural identification.

RESULTSTwo amide alkaloids were obtained. They were identified as N-transferuloyl-3-methyldopamine (I), (10bS)-1,2,3,5,6, 10b-hexahydropyrrolo [2,1-a]-8,9-dihydroxyisoquinoline-3-one (II).

CONCLUSIONCompound I was isolated from this genus for the first time. Compound II is a new compound named salsoline A.

Objective:To valuate the treatment value and analyse the effect on the cellular immune functions by studying the differences of T-lymphocyte subsets and CD4+CD25+Treg cells in peripheral blood after adoptive immunotherapy ( dendritic cells and cytokine-induced killer cells,DC-CIK) combined with chemotherapy on MM.Methods:50 patients with MM were randomly divided into two groups.24 patients in chemotherapy group were treated by chemotherapy only,26 patients in joint group were treated by adoptive immunotherapy( DC-CIK) combined with chemotherapy,and the clinical outcomes and the levels of T-lymphocyte subsets and CD4+CD25+Treg cells in peripheral blood between two groups were compared.Moreover,the differences of cellular immune indicators (Th1/Th2,the ratio of AgNOR,and TGF-β)between two groups were also compared.Results: After treatment,quality of life,clinical index and survival in joint group were better than in chemotherapy group( P<0.05);the proportion of CD3+CD8+,the ratios of CD4+CD25+,CD4+CD25+/CD4+and the level of TGF-βof joint group wes clearly lower than chemotherapy group(P<0.05),and the ratios of CD3+CD4+/CD3+CD8+, Th1/Th2 and AgNOR of joint group wes clearly higher than chemotherapy group .Conclusion: DC-CIK combined with chemotherapy could be an effective and promising treatment to patients with MM,and it maybe strengthen the anti-tumor action of bodies by regulating the balance between Th1 and Th2 reaction.

Objective Calreticulin (CRT)is a multifunctional protein of endoplasmic reticulum implicated in the pathogenesis of rheumatoid arthritis(RA). The present study was undertaken to determine whether CRT was involved in an?giogenesis events in RA. Methods Serum CRT levels were measured by enzyme-linked immnuosorbent assay(ELISA)in 106 patients with established RA, 75 osteoarthritis(OA)and 80 healthy controls(HC). CRT levels in synovial fluid were al?so measured in 25 RA and 22 OA patients. The expression of CRT in synovial tissue was examined by immunohistology. In order to investigate the role of CRT on angiogenesis, human umbilical vein endothelial cells(HUVECs)were isolated and cultured for in vitro experiments. The proliferation, migration and tube formation of HUVECs following CRT stimulation were examined in vitro by MTT assay, scratch wound healing assay and tube formation assay. Results Our results showed a sig?nificantly higher concentration of CRT in serum [(6.4±3.1) μg/L] of RA patients compared to that of OA [(3.7±0.9) μg/L, P<0.01] and HC [(3.4±1.0) μg/L, P<0.01];and significantly higher CRT in synovial fluid [(6.9±3.4) μg/L] of RA vs OA [(3.9± 0.7) μg/L, P<0.01]. Increased CRT expression predominantly localized to vascular endothelial cells, inflammatory cells and perivascular areas in both the lining and sublining layers of RA synovial tissue. Furthermore, CRT significantly promoted the proliferation, migration and tube formation of HUVECs, as showed by MTT assay, scratch wound healing assay and tube for?mation assay. Conclusion These findings suggested that CRT may be involved in synovitis and pannus formation events via promoting angiogenesis in RA.

Objective:To investigate the relationship between the left ventricular morphology, systolic function and the severity of the coronary artery lesions. Methods:A total of 589 consecutively admitted patients were studied retrospectively. All patients suspected of coronary artery disease underwent a selective coronary angiography and a survey of left ventricular ejection fraction (EF) during the hospitalization. Results:A significant correlation between the severity of the coronary artery lesions and the EF, FS, LADD, LADS, LVDS and LVDD was found through the Bivariate Correlation analysis ( P

AIM To explore pharmacokinetics of Berberine in YL2000 in normal and febrile rats. METHODS The levels of Berberine in plasma were measured through HPLC and secondary parameters were obtained by fitting the dose time data of Berberine making use of 3P87 programme. RESULTS In normal and febrile rats, the plasma concentration of Berberine was peaked at (3 4?0 3) h vs (0 3?2 1) h( P

Objective To investigate the effect of extracorporeal shock wave therapy (ESWT) on early and mid-term knee osteoarthritis (KOA). Methods 70 patients were randomly assigned to ESWT group (n=34) or control group (n=36). The ESWT group was given shock wave on unilateral knee, while the control group accepted shock wave with the energy density of 0. They were evaluated with Visual Analog Scale (VAS) on movement, Lequesne Index and Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index before and 12 weeks after the intervention. Results The scores of VAS, Lequesne Index and WOMAC Osteoarthritis Index improved more in the ESWT group than in the control group after intervention (P<0.01). Conclusion ESWT is effective on KOA as a non-surgical treatment.