Objective To translate the findings from fundamental research into clinical application and to evaluate the clinical efficiency and safety of Fritillaria thunbergii Miq. granule as adjunctive means of chemotherapy during peri chemotherapy of refractory acute leukemia. Methods Patients in multiple hospitals were randomly divided into two groups with Fritillaria thunbergii Miq. granule treatment or synchronous control at three days before chemotherapy respectively, according to random approaches for medical treatment. The clinical therapeutic effects were then determined after one course of treatment. Results According to the research project, 138 patients were analyzed statistically, 72 in Fritillaria thunbergii Miq. granule group and 66 in control group. The complete remission rate (CR) of Fritillaria thunbergii Miq. granule group and control group were 36.8% and 25.8% respectively, while the total effects were 77.8% and 53.0%, which was significantly different (P

Objective To explore the syndrome distribution laws of refractory idiopathic thrombocytopenic purpura (RITP), and the relevance between various syndromes and gender, age, laboratory indicators, to reveal the essence of TCM syndrome and provide a reasonable and standard TCM treatment principle. Methods TCM syndrome, gender, age and laboratory indicators of 75 cases of RITP were studied retrospectively and analized statistically. Results Qi deficiency and blood stasis is the major syndrome, and Yin deficiency is the secondary syndrome. Compare with others, the patients with qi deficiency and blood stasis syndrome were older, and had longer disease courses and more severe conditions. Conclusion One of TCM intervention treatment principles for RITP is invigorating qi and yin, promoting blood circulation and removing blood stasis.

Objective To study the impact of compound zhe-bei granule and adriamycin on p53 gene expression in the P388 xenograft tumor.Methods The tumor xenografts model was established by injecting the mouse lymphocytic leukemia cells(P388) in the subcutis of anterior axillary of Kunming mice,and then was treated with CZBG by intragastric administration and different doses of adriamycin by intraperitoneal injection(i.p.).After the end of the experiment,tumor was striped completely,and the expression of p53 gene in xenograft tumor of each group was detected by fluorescence quantitative polymerase chain reaction,and the relative quantification of p53 gene of each group was computed using 2-??Ct method.Results Comparing the 2-??Ct values of p53 gene relative expression of each group,no statistical significance was found(F=0.56,P=0.7557).And the relative expression value of p53 gene of CZBG joint high-dose ADM group was higher,while the relative expression value of CZBG joint low-dose ADM group was lower.Conclusion Combined use of CZBG and ADM is able to raise the expression of p53 tumor suppressor gene in the P388 xenograft tumor.

To investigate the effects of Compound Zhebei Granule (CZBG), a compound traditional Chinese herbal medicine, combined with adriamycin (ADM) on mdr 1 gene expression in tumor xenografts in nude mice.

Objective:To establish an HPLC method with dual wavelength detection for the determination of benzoic acid and cin-namic acid in Suhexiang pills. Methods:A Waters Sun Fire TM C18 column(150 mm × 4. 6 mm,5 μm) was adopted with 0. 1% ace-tic acid-methanol(60∶40) as the mobile phase at a flow rate of 1. 0 ml·min-1 . The detection wavelength was set at 228nm for benzoic acid(0-18min) and 285nm for cinnamic acid (18. 1-35min). The injection volume was 10μl, and the column temperature was room temperature. Results:The linear range of benzoic acid and cinnamic acid was 1. 0-50. 0 μg·ml-1(r=0. 999 7) and 0. 2-10. 0 μg· ml-1, respectively(r=0. 999 9). The average recovery was 96. 88%(RSD=1. 6%, n=9) and 99. 35%(RSD=1. 7%, n=9), re-spectively. Conclusion:The method is accurate, fast and simple in the determination of benzoic acid and cinnamic acid in Suhexiang pills.

The relationship between tumour necrosis factor-alpha (TNF-alpha) gene polymorphism and inhibitory effects of triptolide on TNF-alpha production from peripheral blood mononuclear cells (PBMC) of healthy humans was investigated. Genomic DNA from 41 healthy people was typed for TNF-alpha--308 polymorphism by allele-specific polymorphism chain reaction (AS-PCR). The TNF-alpha concentration in the supernatant was measured by ELISA. The results showed that the production of TNF-alpha from TNF-alpha--308 non-G/G genotype PBMC was higher than that from TNF-alpha--308 G/G genotype PBMC after stimulated by LPS. Triptolide could lower the production of TNF-alpha from G/ G genotype PBMC, but had no effect on the level of TNF-alpha from non-G/G genotype PBMC. It was concluded that TNF-alpha gene polymorphism was related to the TNF-alpha production from triptolide-inhibited PBMC culture in healthy humans.

AIM: To explore whether the inhibitory effect of triptolide on IL - 1β production by PBMC is asso ciated with IL - 1β gene polymorphisms. METHODS: IL - 1β gene polymorphism was analyzed in 31 healthy volunteers. From genomic DNA, the C - T polymorphism at IL - 1 β - 511 was typed by PCR - RFLP. Meanwhile the IL - 1 β was also measured in the supernatants of the cultured and stimulated peripheral blood mononuclear cells (PBMC) by ELISA. RE SULTS: After LPS stimulation in PBMC cultures of healthy subjects, the secretion levels of IL - 1 β in 9 volunteers who carried IL - 1β -511 T/T genotype were higher than in volunteers who are not T/T genotype (P ＜0.05). Triptolide sup pressed the production of IL - 1β significantly in LPS - treated human PBMC carried C/C and C/T genotype ( P ＜ 0.05 ), but this significant inhibitory effect of triptolide was not seen in T/T genotype ( P ＞ 0.05 ). CONCLUSION: The gene polymorphism at IL - 1β - 511 was related to the production of IL - 1β, and the inhibitory effect of triptolide on the produc tion of IL - 1β was different in C/C, C/T, T/T genotype of IL - 1β -511, which may be one of the reasons for the phe nomenon that people respond differently to triptolide.

Objective To evaluate effects of simultaneous inhibition of mammalian target of rapamycin complex 1(mTORC1)kinase and glycogen synthase kinase-3β(GSK-3β)on phosphorylation of 4E-binding protein-1(4EBP1), cap-dependent translation, as well as survival and apoptosis of melanoma cells. Methods Cultured A375 cells were classified into several groups to be treated with dimethyl sulfoxide (DMSO group), the mTORC1 kinase inhibitor everolimus at a concentration of 5 nmol/L (everolimus group), the GSK-3β kinase inhibitor AR-A014418 at a concentration of 10 μmol/L (AR-A014418 group), or 5 nmol/L everolimus and 10 μmol/L AR-A014418(combined treatment group). After additional culture, Western-blot analysis was performed to measure protein expressions of phosphorylated 4EBP1 (p4EBP1)and survivin in A375 cells, m7GTP pull down assay to estimate interaction between eukaryotic initiation factor-4E (eIF4E)and eIF4G, cell counting kit 8 (CCK8)assay to evaluate cell proliferation, and flow cytometry to detect cell apoptosis. Results Both everolimus and AR-A014418 had inhibitory effects on 4EBP1 phosphorylation and survivin expression. The expressions of p4EBP1-65 and survivin were both significantly decreased in the everolimus group (0.74 ± 0.05 and 0.71 ± 0.06 respectively), AR-A014418 group (0.62 ± 0.06 and 0.58 ± 0.07 respectively)and combined treatment group (0.14 ± 0.04 and 0.09 ± 0.05 respectively)compared with the DMSO group (1.00 ± 0.07 and 1.00 ± 0.06, respectively, all P < 0.001), with the most significant decrease observed in the combined treatment group. As m7GTP pull-down assay showed, the everolimus group, AR-A014418 group and combined treatment group all showed significantly lower relative expression levels of eIF4G(0.72 ± 0.04, 0.67 ± 0.05 and 0.12 ± 0.05 vs. 1.00 ± 0.06, all P < 0.001), but significantly higher relative expression levels of 4EBP1 (1.98 ± 0.16, 2.32 ± 0.17 and 7.58 ± 0.25 vs. 1.00 ± 0.08, all P < 0.001)than the DMSO group, and the combined treatment group showed the lowest eIF4G expression but highest 4EBP1 expression. After 24-hour culture, the proliferation of A375 cells was inhibited by 18.5% ± 1.3% in the everolimus group, 19.8% ± 1.8% in the AR-A014418 group, and 61.2% ± 2.1% in the combined treatment group compared with the DMSO group, with the strongest inhibition noted in the combined treatment group. The inhibitory effects of everolimus and AR-A014418 on cell proliferation increased over time, and showed the same trend at 48 hours. Flow cytometry showed that the apoptosis of A375 cells was accelerated by the 24-hour treatments with everolimus and AR-A014418 alone or in combination, with the apoptosis rate being 14.28% ± 2.18%, 14.57% ± 2.35% and 55.18% ± 6.27% in the everolimus group, AR-A014418 group and combined treatment group respectively, and the combined treatment showed the strongest accelerating effect. Conclusion The combined treatment with everolimus and AR-A014418 can evidently inhibit 4EBP1 phosphorylation and eIF4F complex formation in A375 cells, which then suppress cap-dependent translation and promote apoptosis of melanoma cells.

Objective:To investigate the expression of microRNA-Let-7a in the serum and tumor tissue of non-small cell lung cancer( NSCLC) patients and the effects of cancer cell migration and proliferation.Methods: 50 cases of NSCLC patients in our hospital as the study group,50 healthy volunteers were used as control group,we used Real-time RCR to detect the expression of microRNA-Let-7a in the serum and tumor tissue of NSCLC patients.Using microRNA let-7a mimics transfected into A549,the level of cancer cell migration was observed by transwell,the level of cell proliferation was observed by CCK-8,the level of k-Ras was observed by Real-time RCR and Western blot.Results: The expression of microRNA-Let-7a in the serum and tumor tissue of NSCLC patients was significantly higher than the control group,the difference was statistically significant (P<0.05).After microRNA let-7a transfected into A549,the levels of cancer cell migration and proliferation were significantly decreased,the difference was statistically significant (P<0.05),the mRNA and protein levels of k-Ras were reduced inA549 cells,the difference was statistically significant (P<0.05). Conclusion:The expression of microRNA let-7a is low in the serum and tumor tissue of NSCLC patients,and may weaken the levels of cancer cell migration and proliferation through the Ras signaling pathway.

Objective To investigate the clinical characteristics and high-risk factors of parenteral nutrition-associated cholestasis (PNAC) in very low birth weight infants (VLBWI),so as to guide the prevention and treatment of this condition.Methods A retrospective analysis was conducted with the data of 141 VLBWI collected from January 2013 to October 2014 in Affiliated Hospital of Qingdao University.These VLBWI were of gestational age ≤ 34 weeks and received parenteral nutrition (PN) for ＞ 14 days in Neonatal Intensive Care Unit.According to the existence of PNAC,they were divided into PNAC group (n =32) and non-PNAC group (n =109) and analyzed in terms of clinical features.Results Between the non-PNAC and the PNAC groups,univariate analysis showed significant differences in gestational age [(30.84 ± 1.55) weeks vs.(29.68 ±2.04) weeks,t=2.952,P=0.005],birth weight [(1 267.92±160.39) gvs.(1 050.63 ±229.74) g,t=6.064,P=0.000],hospital stay [(43.26 ±14.43) d vs.(66.47 ±22.25) d,t =-6.622,P=0.000],fasting time [(6.30±5.23) d vs.(11.94±7.92) d,t＝-4.723,P=0.000],PN duration [(32.40 ± 13.72) d vs.(57.59 ± 27.65) d,t =-7.039,P =0.000],the maximum daily dosage of amino acid [(3.61 ±0.27) g/ (kg·d) vs.(3.78±0.35) g/ (kg·d),t=-3.012,P=0.003];concurrent infection (56.88％ vs.87.50％,x2 =10.046,P =0.002),pneumonia (32.11％ vs.68.75％,x2 =13.790,P =0.000),necrotizing enterocolitis (0.92％ vs.9.38％,x2 =6.420,P =0.037),neonatal respiratory distress syndrome (55.05％ vs.78.13％,x2 =5.503,P =0.019),bronchopulmonary dysplasia (19.27％ vs.46.88％,x2 =9.918,P=0.002),congenital heart disease (4.59％ vs.21.88％,x2 =7.405,P =0.007);application of pulmonary surfactant (33.94％ vs.59.38％,x2 =6.682,P =0.010),continuous positive airway pressure ventilation (60.55％ vs.87.50％,x2 =8.085,P =0.004),mechanical ventilation (22.94％ vs.43.75％,x2 =5.356,P =0.021) and peripherally inserted central catheter (44.04％ vs.81.25％,x2 =13.737,P =0.000).Multivariate Logistic regression analysis revealed that PN duration was an independent high-risk factor of PNAC (B =0.069,OR =1.071,95％ CI =1.039-1.104,P =0.000).Conclusions Many factors are associated with PNAC,in which PN duration is an independent high-risk factor.Early enteral nutrition and shortened duration of PN may be protective measures for PNAC.