Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People’s Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories’ results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.

OBJECTIVE： To observe improvement effects of fingolimod on middle cerebral artery occlusion/reperfusion （MCAO/R） injury model rats. METHODS： Male SD rats were randomly divided into sham operation group， model group and fingolimod low-dose， medium-dose and high-dose groups （0.5， 1， 2 mg/kg）， with 8 rats in each group. Except for sham operation group， MCAO/R injury model was induced by suture-occluded method in other groups. Administration groups were given relevant medicine intragastrically after reperfusion [1 h after reperfusion （1st day）， 22.5 h after reperfusion （2nd day）， and then every 24 h until 142.5 h of reperfusion （7th day）]. Sham operation group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 7 d. The scores of neurological deficit and balance beam test， the times of memory error [work memory error （WME）， reference memory error （RME） and total error] were recorded in each group. The contents of serum inflammatory cytokines （IL-6， IL-8， IL-10， TNF-α） were determined by ELISA， and triphenyl tetrazolium chloride staining method was used to detect the rate of cerebral infarction. RESULTS： Compared with sham operation group， neurological deficit scores （at different time points of 1st-7th day after administration）， balance beam test scores （2nd， 4th， 7th day after administration）， times of memory error （2nd， 4th， 7th day after administration）， the contents of serum inflammatory cytokines and the rate of cerebral infarction were increased significantly in model group （P＜0.05 or P＜0.01）. Compared with model group， neurological deficit scores （low-dose group at different time points of 3rd-7th day， medium-dose and high-dose groups at different time points of 2nd-7th day after administration）， balance beam test scores （low-dose group at 7th day， medium-dose group at 4th and 7th day， high-dose group at 2nd， 4th， 7th day）， RME times and total error times （low-dose group at 4th and 7th day， medium-dose group and high-dose group at 2nd， 4th， 7th day after administration）， WME times （administrations groups at 7th day after administration）， serum contents of IL-6， IL-8 and IL-10 （administrations groups）， serum contents of TNF-α （medium-dose and high-does groups） and cerebral infarction rate （medium-dose and high-dose groups） were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Intragastric administration of fingolimod can significantly reduce neurological deficit score， balance beam test score and the times of memory error in MCAO/R injury model rats， and has a protective effect on cerebral tissue and memory function. These effects may be related to the down-regulation of inflammatory cytokines such as IL-6 and TNF-α by fingolimod.

This paper reviews the history of traditional Chinese medicine powder from germination, birth, prosperity to the clinical application, which is gradually reduced. And it enumerates the clinical application of traditional Chinese medicine powder taken orally, external application. The powder preparation process are outlined, summarizes the preparation results including crushing, drying, mixing, taste masking and inhibition of volatilization, sterilization with combining innovation and advice of researchers in the process of powder research. It discussed the main problems of restricting large-scale production that running through preparation, quality standard, clinical application (such as dependence of patients) of powder. Then, it forecasted that more and more hospitals and families will use traditional Chinese medicine powder to relieve pain of patients, in order to enhance the level of preparation and quality control, boosting the normalization and standardization of powder.

Objective To evaluate the efficacy and safety of Houpu Paiqi mixture in treatment of functional dyspepsia (FD)with abdominal distension symptom.Methods From July 2014 to June 2015 , in nine centers,a total of 162 FD patients with abdominal distension symptom and met Rome Ⅲpostprandial distress syndrome (PDS)diagnostic criteria were enrolled.All patients were randomly divided into trial group and control group,81 patients in either group.The patients of trial group and control group took Houpu Paiqi mixture or placebo,respectively,25 mL per time,twice daily,and both the courses of treatment were two weeks.Before and after the treatment,the improvement of main symptoms,total clinical efficacy rate and efficacy of traditional medicine between two groups were compared.Chi square test,Fisher exact probability method and Wilcoxon test were performed for statistical analysis.Results According to the results of per-protocol (PP)analysis,the total efficacy rate of trial group and control group was 69.4% (50/72)and 59.2% (42/71),respectively,and there was no statistically significant difference in total efficacy rate between the two groups (χ2 =1 .650,P =0.199 ). And there was no statistically significant difference in the improvement of PDS main symptoms(postprandial fullnessand early satiety)between the two groups (56.3% ±27.9% vs 54.4% ±32.1%,t =0.606,P =0.727 ).For those with baseline symptom score over 14,median early satiety score of trial group after the treatment was 0,which was lower than that of control group,and the difference was statistically significant (Z =-2.370,P =0.018).The total efficacy rate of traditional medicine of trial group was 80.8% (59/73 )and that of control group was 72.0% (54/75 ),and the difference was not statistically significant (χ2 = 0.676,P =0.411 ).Conclusion Houpu Paiqi mixture has certain efficacy in FD with abdominal distension,and could be used for the treatment of PDS-predominant FD.

Objective To explore the influence of core competence evaluation on continuing education of training nurses in operation room.Methods Questionnaires were administered to 42 training nurses by using the registered nurse core competence scale (CIRN) in operation room.Results The level of core competence of training nurses in operation room was medium (3.25 ± 0.45).Legal and ethical practice dimension of scores was the highest (3.55 ± 0.63).The single average of clinical nursing dimension was the advanced lowest (3.11 ± 0.54).The nurses self identity was the core influencing factors by multiple stepwise regression analysis in regression.Conclusions The core competence of training nurses in operation room is on the middle level,The focus is to improve the ability of clinical nursing in continuing education.To give more attention and support in self identity of the operation room training nurses,and to provide the scientific basis for the training plan and specialist training for them,in order to promote the continuing education projects of training nurses.

Objective To investigate the mechanism of P/Q-type calcium channel in nitroglycerin induced migraine experimental model Methods Twenty Sprague-Dawley rats ( half female and half male) were randomly divided into control group and model group. The model of migraine rats was reproduced using Tasserelli Cristina method that wassubcutaneou injection of GTN of 10 mg/kg, once a week for four weeks. After the model of migraine had been established, trigeminal ganglion and trigeminocervical complex and cortex of frontal lobe were removed and the expressions of P/Q-type calcium channel were detected by RT- PCR and Western-blot, and at the same time the concentration of intracellular Ca2+ was investigated by Fluo- 3/AM fluorometric method. Results Compared with the control group, the expression of mRNA and protein of P/Q-type calcium channel of trigeminal ganglion and trigeminocervical complex (mRNA: 0. 472 36± 0. 049 54; protein: 0. 337 25 ± 0. 035 93 ) and cortex of frontal lobe ( mRNA: 0. 547 45 ± 0. 044 39 ; protein : 0. 402 13 ± 0. 029 83) in model group all upregulated (t = 2. 6697, 3. 1993, 3. 4398 and 3. 7661, all P <0. 05), at the same time the concentration of intracellular Ca2+ in model group increased (211 182 ± 12 973 vs 135 243 ± 18 105 in trigeminal ganglion and trigeminocervical complex; 186 511 ± 18 297 vs 143 289±25 175 in cortex of frontal lobe. t =10.7819 and 4. 3917, beth P<0.05). Conclusions P/Q- type calcium channel may play a role in the pathogenesis of migraine via its upregulated expression.

The LACK gene from Leishmania, an analogue of the receptor of activated protein kinase C, was discovered recently. In this study, the LACK gene of Leishmania donovani was obtained from the recombinant plasmid T-LACK by PCR. The gene was cloned into eukaryotic expressed plasmid pcDNA3.1(+) to construct recombinant plasmid. This recombinant plasmid then was transfected into the eukaryotic cell COS-7, and the expression of LACK gene in eukaryotic cell was detected by RT-PCR and immunofluorescent staining. Both RT-PCR and immunofluorescent staining of recombinant plasmid transfected COS-7 showed positive reaction, thus indicating that the recombinant plasmid pcDNA3-LACK can express LACK protein in euka ryotic cell COS-7.
Animals ; Antigens, Protozoan ; biosynthesis ; genetics ; immunology ; COS Cells ; Cloning, Molecular ; DNA, Recombinant ; biosynthesis ; genetics ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Leishmania donovani ; Plasmids ; genetics ; Protozoan Proteins ; biosynthesis ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Vaccines, DNA

OBJECTIVETo confirm the existence of point mutations in the SSU rDNA variable regions of 5 Leishmania donovani (L.d.) isolates from different epidemic foci in China.

METHODSSpecific SSU rDNA fragments from nuclear DNA of 7 Leishmania species/isolates were amplified by PCR and then cloned into pGEM(R)-T Easy Vectors. After that, the specific fragments were sequenced by an automated DNA sequencer.

RESULTSSequence analysis showed that the amplified DNA fragments of 7 Leishmania species/isolates were all 392 bp in length. All 5 point mutations were located in two unique sequence blocks (UQ-I and UQ-II), and no insertions or deletions were found. The identities of comparison of Leishmania in GeneBank were more than 98%.

CONCLUSIONFive point mutations exist in the SSU rDNA variable region of 5 L.d. isolates from different epidemic foci of visceral leishmaniasis (VL) in China. Sequence differences of the SSU rDNA variable region exist among L.d. isolates from different foci.

Animals ; DNA, Protozoan ; chemistry ; DNA, Ribosomal ; chemistry ; Humans ; Leishmania donovani ; genetics ; Leishmaniasis, Visceral ; parasitology ; Point Mutation ; Polymerase Chain Reaction

Objective] By sequencing of SSU rRNA gene cloning from Xinjiang cutaneous leishmaniasis pathogen (XJCLP) to provide evidence for identification of the pathogen. [Methods] By PCR assay with primers R222 and R333, the specific fragment had been produced from SSU rRNA gene of XJCLP , L infantum, L tropica and cloned into pGEM ○[KG-6/7]R T Easy vector .The clones were sequenced by the Sanger dideoxy mediated chain termination method, analysis of SSU rRNA gene sequences from XJCLP, L tropica, L infantum with DNASIS. [Results] Sequence analysis showed that the specific fragment of SSU rRNAgene from XJCLP, L infantum,L tropica , were all 394 bp in length. There were 391 bases identical and three point mutations between the sequences of XJCLP and L tropica , the similarity being 99 2%; 390 bases identical and three point mutations and one insertion /deletion between the sequences of XJCLP and L infantum , the similarity being 99 0%. One insertion/deletion between the sequences of L tropica and L infantum , the similarity being 99 7%. The primary and secondary structures of SSU rRNA gene from XJCLP differed from those of L infantum and L tropica .A retrieval from GenBank confirmed that these 394 bp sequence are new gene sequences. [Conclusion]The primary and secondary structures of SSU rRNA gene from XJCLP, L infantum , L tropica were different. 394 bp sequence from SSU rRNA gene of XJCLP is a new gene sequence.

Objective] To analyze the sequence difference of the SSU rDNA variable regions of Leishmania isolates from hilly foci and plain foci of China. [Methods] Specific SSU rDNA fragments from nuclear DNA of five Leishmania species and isolates were amplified by PCR. The amplified DNA fragments were cloned into pGEM R\|T Easy vector. The specific fragments were sequenced by the automated DNA sequencer. [Results] Sequence analysis showed that the amplified DNA fragments of five Leishmania species and isolates were all 392 bp in length, point mutations were located in the two unique sequence (UQ\|Ⅰ and UQ\|Ⅱ); L.d.SC10 and L.d.GS7 had two same point mutations in UQ\|Ⅱ, only L.d.GS7 had one in UQ\|Ⅰ; no insertion/deletion. [Conclusion] Sequence difference of the SSU rDNA variable region existed between Leishmania isolates from hilly foci and plain foci; The sequences of the SSU rDNA variable regions of L.d. SD2 isolate and L.infantum were identical.