Objective:To detect A20 mutation and to investigate its relationship with clinicopathologic features, prognosis, and re-sistance to therapy of diffuse large B cell lymphoma (DLBCL). Methods:A total of 104 cases with DLBCL and their clinical data were collected;follow-up was performed on a few of DLBCL patients. The expression of P-gP and Ki-67 protein was detected with immuno-histochemical staining. The A20 gene mutation in exons 3, 6, and 7 were examined by polymerase chain reaction and DNA sequencing. Finally, the correlation of genetic abnormality of A20 with clinicopathologic features was analyzed. Results:Missense mutation in ex-on 3 of A20 gene was identified in 18 of 104 patients (17.3%). The A20 gene mutation at site 73 of exon 3 was greater in the cases with activated B cell-like-DLBCL than with germinal center B-cell-like-DLBCL (18.5%vs. 2.5%, P=0.010). In contrast to the advanced clin-ical stage and high International Prognostic Index (IPI) cases, the rate of A20 mutation was superior to the opposite (P<0.05). The ex-pression for P-gP was higher in the cases with mutation than that of those with wild-type A20 gene (16/18 vs. 52/86, P=0.021). The dif-ference in A20 mutation between the groups of low and high positive expression for Ki-67 (1/17 vs. 26/60, P=0.030) was significant. DLBCL with A20 mutation had an increasing recurrence tendency (P=0.06) and a worse survival (P=0.016) compared with those with wild-type A20 gene. Conclusion:The A20 mutation has a certain influence on the clinicopathologic characteristics and survival condi-tions of BLBCL patients. Probably, A20 mutation is a factor associated with a poor prognosis of DLBCL.

Objective To study the changes of chemical constituents and pharmacodynamics with different drying methods (sun- drying, 50 ℃ vacuum-drying, 50 ℃ and 80 ℃ heat-drying, freeze drying method) in Bufonis Venenum. Methods HPLC method and TLC method was established for studying the changes of chemical constituents from B. Venenum before and after being dried, and determine the inhibitory effect of dried samples on five different tumor cell lines proliferation by MTT assay. Results The B. Venenum processed by five different methods were different in character, while no significant differences in the types and content of chemical constituents; The total content of resibufogenin and cinobufagin was more than 6%, which was consistent with the 2015 edition of Chinese Pharmacopeia; 50 ℃ and 80 ℃ heat-drying of B. Venenum showed more effective inhibitiory effect than other dry methods. Conclusion The appearance of B. Venenum met the criterion of pharmacoperia by sun-drying and 50 ℃ heat-drying method. Although the color of B. Venenum under the vacuum-drying and freeze drying method did not meet the requirements of Chinese Pharmacopeia, the main effective components have a few changes.

OBJECTIVETo construct and identify a lentiviral vector of RNA interference targeting human Notch-1 gene.

METHODSTo determine the Notch-1 gene sequences, three RNAi target sequences (shRNA1-3) were designed in accordance with the RNAi sequence design principles and cloned into the lentiviral vector pLenOR-THM by endonuclease BamH I restriction, EcoR I double digestion, and T4 DNA-ligase ligation. After the transformation into competent DH5alpha bacteria, the candidate clones were identified by Kpn I and EcoR I double digestion and DNA sequencing. The recombinant and three packaging plasmids were co-transfected into human embryonic kidney cell line 293T cells by lipofectamine to produce the lentiviral particles. The viral titer was determined. The 293T cells were infected by the lentiviral particles obtained, and transfection efficiency was assessed using a fluorescent microscope. The lentiviral vector particles were also transfected into ACC-M cells. The Notch-1 expression in the transfected cells was assayed by quantitative reverse transcription polymerase chain reaction (QRT-PCR) and Western blot analysis.

RESULTSThe lentiviral RNAi vector pLenOR-THM-Notchl for Notch-1 gene was constructed successfully. Strong green fluorescence was observed in the 293T cells under fluorescent microscope after co-transfection of the cells with the four plasmids of lentiviral vector. The virus in the supernatant reached a titer of 5.8 x 10(8) TU x mL(-1). The transfection efficiency of the collected virus exceeded 90% in 293T cells with 1 as a multiplicity of infection. The third lentiviral vector was found to significantly inhibit the Notch-1 expression at the mRNA and protein levels.

CONCLUSIONThe lentiviral RNAi vector of Notch-1 has been successfully constructed and identified.

Cell Line ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Notch1 ; Transfection

This paper introduced the application and effects of mobile operating room nursing information management system (MORNIMS) in the operating room. The MORNIMS was designed by operating room and information center based on the Enterprises Digital Assistant (EDA) and was applied in the operating room for two years. Its application significantly improved work efficiency,optimized the work flow,reduced nursing mistakes,enhanced the retrospective quality monitoring of sterile materials,and realized information management in the operating room.

This study was aimed to screen out a safe and effective traditional Chinese medicine (TCM) compound prescription in insomnia treatment, through the method of uniform design and comprehensive scoring method. It provided candidate drugs to further research and development of TCM new drug. U7(76) uniform design was used to divide groups. The test was given on sleep time prolonging effect of mice injected with pentobarbital sodium. The subliminal dosage of pentobarbital hypnosis was tested on mice injected with pentobarbital sodium. The acute toxicity (LD50) was tested on mice. Comprehensive scoring method was applied to evaluate the effect to determine the optimum dose. The results showed the optimized prescription was composed of Radix Sophorae Flavescentis and Schisandra chinensis. The optimum ratio was 1:3. It was concluded that the uniform design was applied in the experimental design with 3 factors and 6 levels. Comprehensive scoring method was used to evaluate the combined effect. The optimized prescription that put benefit and risk into consideration was composed of Radix Sophorae Flavescentis and Schisandra chinensis. The best ratio was 1:3, which can be further developed.

AIM: To explore whether necroptosis contributes to the high glucose (HG)-induced damage in hu-man umbilical vein endothelial cells (HUVECs).METHODS: The protein levels of receptor-interacting protein 3 (RIP3) and cleaved caspase-3 were detected by Western blot.The intracellular levels of reactive oxygen species (ROS) were deter-mined by DCFH-DA staining followed by photofluorography.Mitochondrial membrane potential (MMP) was measured by rhodamine 123 staining followed by photofluorography.RESULTS: Treatment of HUVECs with HG at different concentra-tions (10, 20 and 40 mmol/L glucose) for 24 h gradually enhanced the expression levels of RIP3.Treatment of HUVECs with HG (40 mmol/L glucose) for different time (3 h, 6 h, 9 h, 12 h and 24 h) also up-regulated the expression levels of RIP3, peaking at 9 h.Pretreatment of HUVECs with 20 μmol/L Z-VAD-FMK (an inhibitor of caspase) for 30 min before exposure to HG enhanced the expression level of RIP3.Pretreatment of HUVECs with 100 μmol/L necrostatin-1 (an inhi-bitor of necroptosis) for 1 h before exposure to HG alleviated the HG-induced injuries, such as a decrease in cell viability, an increase in ROS generation and dissipation of MMP, but up-regulated the protein level of cleaved caspase-3.CON-
CLUSION: Necroptosis mediates HG-induced injury in HUVECs.There is a negative interacting between necroptosis and apoptosis.

To explore the effects of recombined Neuregulin on the heart of healthy Macaca mulatta, 10 healthy adult Macaca mulatta were randomly divided into two groups and were injected with the same doses of recombined Neuregulin and normal saline, respectively. At the same time, related indices were detected by 2-dimensional echocardiography and M-mode echocardiography. All indices were compared between the two groups and among different phases. Recombined Neuregulin had effects on LVEDD (left ventricular end-diastolic diameter), LVEDV (left ventricular end-diastolic volume), LVESV (left ventricular end-systolic volume) and SV (Stroke volume), and the effects changed with time. However, no significant changes were seen on EF (Ejection fraction) and FS (Fractional shortening). In conclusion, recombined Neuregulin has effects on the left ventricular volume of healthy Macaca mulatta, but no significant effect on cardiac contractility.
Animals ; Echocardiography ; methods ; Female ; Heart Ventricles ; anatomy & histology ; diagnostic imaging ; drug effects ; Macaca mulatta ; Male ; Myocardial Contraction ; drug effects ; Neuregulins ; biosynthesis ; genetics ; pharmacology ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Stroke Volume ; drug effects

BACKGROUNDTo compare the therapeutic effect, adverse reaction and effect on immunity of chemotherapy combined Aidi injection (AI) with those of chemotherapy alone in the treatment of advanced non small cell lung cancer (NSCLC).

METHODSNinety eight cases of advanced NSCLC were randomly divided into two groups, trial group and control group. In the trial group, NP plus AI (60 80 ml) were given intravenously by dissolving in 400 ml of normal saline per day for 8-10 days, while in the control group, only NP chemotherapy was given. Navelbine (25 mg/m², d1, 8) and cisplastin (40 mg/m², d1-3) were chosen in the chemotherapy. Each patient received at least two cycles of treatment.

RESULTSThe effective rate in the trial group and the control group was 53.1% and 44.9% respectively, without significant difference between the two groups ( P > 0.05). But the rate of progression, adverse reactions in bone marrow and digestive tract, and change of immunity in the trial group were all lower than those in the control group ( P < 0.05), and the improvement in Karnofsky score in the trial group was higher than that in the control group ( P < 0.05).

CONCLUSIONSChemotherapy of NP combined with AI shows benefit in the treatment of advanced NSCLC. AI could decrease the influence on immunity and adverse reaction of chemotherapy, and improve the quality of life in patients with NSCLC.

【Objective】 To verificate the performance of enzyme-linked immunosorbent assay (ELISA) in blood screening laboratory. 【Methods】 The repeatability, precision, sensitivity, specificity, compliance, detection limit and anti-interference of ELISA items in the laboratory detection system were verified. 【Results】 The repeatability was 100%.The intra batch imprecision of each system was less than 10%, and the inter batch imprecision was less than 15%. The sensitivity, specificity and compliance were 100%, with the minimum detection limits of the two reagents at 0.75 NCU/mL and 0.25 NCU/mL respectively, The anti-interference met the requirements of the reagent manual. 【Conclusion】 The analysis of the performance verification data of ELISA test items will help continuously improve the performance of detection system and ensure the safety of clinical blood use.

To develop an intervention protocol for children's unintentional injury risk behaviors, and to evaluate the feasibility of the protocol.
 Methods: By theoretically analyzing the influential factors for children's unintentional injury risk behaviors, children's cognitive development characteristics and the social learning theory, an intervention protocol was established on the basis of changing the unintentional injury attribution and negative information transmission of risk behavior consequences. A primary school in Changsha city was selected by random cluster sampling. A community-based randomized controlled trial was conducted on the selected students once a week for 5 consecutive weeks. The scores of unintentional injury risk behavior before intervention, 3 months and 6 months after intervention, and the frequency before intervention and 6 months after intervention, were collected and compared.
 Results: A total of 194 children were included in the study: 98 in the intervention group; 96 in the control group; 96 (49.5%) boys and 98 (50.5%) girls between 7 and 8 years old. The scores of unintentional injury risk behavior for children in the intervention group at 3 and 6 months after intervention were 14.42±5.67 and 14.14±8.95, respectively, lower than those before the intervention (16.85±8.48) and in the control group (P=0.001). The number of minor unintentional injuries in the intervention group decreased from 119 to 56, and the number of children suffering 2 or more injuries dropped from 34 to 10 (P<0.001) at 6 months after the intervention, while both of them were lower than that in the control group (P=0.011). Similar changes were observed in some slight or more serious unintentional injuries (P=0.030).
 Conclusion: The protocol for changing the attribution to unintentional injury and negative information transmission for risk behavior consequences was proved to effectively reduce children's unintentional injury risk behaviors and relevant events.
Child ; China ; Cognition ; Female ; Humans ; Male ; Risk-Taking ; Schools ; Students ; Wounds and Injuries ; prevention & control