Objective To analyze genetic relationship of Leishmania species and strains from China by RAPD technique. Methods DNAs from Leishmania strains, including L donovani(L d.) isolates from patients, dogs and sandflies of three different foci in China and international reference strains, were amplified by seven random primers. The DNA polymorphic bands detected were analyzed by clustering analysis with SPSS software. Results ① L d . isolates from hill and plain foci in China were divided into two groups. The genetic distance of L d .isolates is distant between them. ② L d .XJ771, L d .XJ901, L d .XJ801 from desert, vicinity of desert, and plain regions in Xinjiang were in the same group.It indicated that the genetic distance among L d .isolates from the three regions is close. ③ L d .isolated from VL patients and dogs in hill foci could not be discriminated distintly, showing high homology between them. ④ L d .DD8 from India,the reference strain of plain type,was clustered with L d .isolates from plain foci in China.It provided scientific basis for the viewpoint "Kala Azar from east area of China is similar to that from India". ⑤ L infantum and L d .isolates from hill foci in China were clustered into different groups. ⑥ The genetic distance is close between L d .isolates from plain foci in China and L d .Jed;⑦ L infantum and L tropica showed the closest genetic distance. Conclusion Differences at genetic level exist in Leishmania isolates from different foci in China.

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] 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 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.

OBJECTIVE Researching the dynamic changes of air bacteria content in the process of spine operating.METHODS Adopt settling process to monitor the bacterium content of the air in the process of surgery operating at the point of pre-operating,cut skin,surgical sutures,suturescontent.RESULTS There are two obvious peak values of bacteria content in the process of spinal surgery operating,at beginning of operating and operating finished.CONCLUSIONS There are two obvious peak values of bacteria total amount changes in the process of operating in operating room,the main factor of the bacteria total amount changes in the air is the personal activities in operating room during operatings.

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.

Objective To determine the nucleotide sequence of the LACK (Leishmania homologue of receptors for activated protein kinase C) gene of Leishmania donovani isolates from plain foci (L.d SD1) and desert foci (L.d XJ771) of China, and to find out the difference of the sequence of LACK gene with other Leishimania spp. and also the isolate from hill foci of China.. Methods. The LACK genes of L.d SD1 and L.d XJ771 were amplified by RT-PCR and cloned into pUC18 vector respectively, sequenced by the dideoxy chain termination method, then analyzed and compared with that of other isolates.. Results . The LACK genes of the two isolates were successfully cloned. Both of the 2 fragments were 942 bp in length. Comparison of the two nucleotide sequences with that of other Leishmania spp. in GenBank showed that the identities were more than 97%, and the identities of the nucleotide sequences of LACK genes of the three L.d isolates from plain, desert and hill foci of China were more than 95%.. Conclusion . High identities exist among the nucleotide sequences of LACK genes of the three L.d isolates from the three foci of China.

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 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.