Objective To improve the accuracy of detection through analyzing the phenotypes of P.pneumotropica isolates in laboratory animals in Beijing area .Methods 306 suspicious P.pneumotropica strains were identified by biochemical identification and 16S rDNA sequencing.Then, to obtain the phylogenetic relationships combined with colony characteristics on blood agar plates and biochemical characteristics of 53 biotypes .Results BD Phoenix 100 automated bacterial identification system and 16S rDNA sequencing identified P.pneumotropica positive rate of 306 isolates were 164/306 and 227/306, respectively.There were 140 phenotypes in 227 true-positive strains, of which 106 were biotype Heyl and 23 were biotype Jawetz .Conclusions In the samples of laboratory animals in Beijing area , P.pneumotropica infection mainly are of biotype Heyl , and less is of biotype Jawetz .The phenotypes are diverse and widely distributed .

Objective To develop a TaqMan MGB probe-based,sensitive and specific real-time fluorescence quantitative PCR assay for rapid detection of Helicobacter hepaticus.Methods Primers and probes specific toflaB gene of Helicobacter hepaticus were designed.A TaqMan MGB probe-based,real-time fluorescence quantitative PCR was established.The specificity,sensitivity and stability of the assay were assassed.Then,the established TaqMan MGB probe real-time fluorescence quantitative PCR assay was applied to detect Helicobacter hepaticus in 1081 clinical specimens during 2008-2011,compared with bacterial isolation and culture method and conventional PCR assay.Results The specificity of this established TaqMan MGB probe-based real-time fluorescence quantitative PCR was high and there were no cross-reactivity with Helicobacter pylori,Campylobacter jejuni,Clostridium piliforme,Pasteurella pneumotropica,Escherichia coli,Pseudomonas aeruginosa.The detection limits was 8.3 copies.The correlation coefficient and slope value of standard curve were 0.999 and -3.227,respectively and the efficiency of TaqMan MGB-based probe realtime fluorescence quantitative PCR assay was 100％.The TaqMan MGB-based probe real-time fluorescence quantitative PCR and conventional PCR were preformed to detect Helicobacter hepaticus in 1081 clinical specimens,a total of 86 specimens were positive for Helicobacter hepaticus.However,there was only 4 specimens were positive by bacteria isolation and culture method.The results showed that TaqMan MGB -based probe real-time fluorescence quantitative PCR for Helicobacter hepaticas was more sensitive than bacteria isolation and culture method,and it could detect Helicobacter hepaticus DNA from clinical specimens directly,and detection time is only 2 hours.Conclusion The TaqMan MGB-based probe real-time fluorescence quantitative PCR assay was a reliable,specific,sensitive and useful tool for rapid detection of Helicobacter hepaticus.

Objective:we aim to determine the relationship between Cell sarcoma (c-Src) expression in patients with EOC and the disease phenotype.Methods:c-Src expression was evaluated using Western blotting analysis in 21 ovarian carcinomas and 4 normal ovarian tissues.Immunohistochemistry was used to evaluate c-Src expression in 134 ovarian carcinomas and 26 normal ovarian tissues.The association between c-Src expression and clinically pathologic characteristics were also assessed in these patients.Results:Our results indicated elevated c-Src protein in EOCs compared with that in normal tissues.The overexpression of c-Src was significantly associated with aggressive features,such as advanced disease stage,poor histological grade,lymph node metastasis,and tumor recurrence (P＜0.05).In addition,the overexpression ofc-Src is significantly associated with EOCs' prognosis.Conclusion:c-Src overexpression was significantly associated with the malignant biological behavior of tumor,suggesting c-Src as a potential preventive target in these patients.

Objective To establish an effective PCR assay for leptospirosis detection , and applicate the assay in tree shrew, mongolian gerbil and gray hamster .Methods Sequence of leptospira was obtained from the NCBI Genbank , and primers were designed based on the sequences .The positive amplified fragments were sequenced to verify the reliability of the method.The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method .Results The PCR method for detection of leptospirosis was successfully established .The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews , 100% in 104 samples of the conventional Mongolian gerbils , and 0% in 60 samples of clean gray hamsters.Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster .The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research .

Objective To develop a multiplex polymerase chain reaction ( mPCR) assay for detection of four pathogenic dermatophytes [Trichophyton mentagrophytes (Tm), Microsporum gypseum (Mg), Microsporum canis (Mc), and Arthroderma simii ( As) ] in laboratory animals, which could be used rapidly and simultaneously for direct detection of those four pathogens.Methods We designed 5 specific primers according to 18S-28S rRNA sequences of the four pathogenic dermatophytes reported in Genbank. The four mPCR assays were established through optimizing the concentration of primers, dNTP, TaqDNA polymerase and the annealing temperature.After verifying the specificity and sensibility, this method was used to detect 15 hair samples with artificial infection and 260 samples taken from laboratory animals.Results This mPCR technique can distinguish the four dermatophytes by producing 192 bp( Tm) ,460 bp( Mg) , 290 bp( Mc) and 602 bp( As) fragments.The sensibility for detection of the four dermatophytes was 5.9 pg/μL, 6.6 pg/μL, 9.5 pg/μL and 5.1 pg/μL, respectively.The results of 15 artificial infection samples were accurate, and the results of 260 hairs samples were negative for the four fungi.Conclusions Our results suggest that the mPCR assay developed in this study can efficiently detect the four dermatophytes, is a useful and rapid technique for rapid detection of the pathogenic dermatophytes in laboratory animals.

Objective To explore the effect of electromyogram-triggered neuromusclar stimulation (ETNS) on motor function of upper limbs of stroke patients. Methods 45 stroke patients from July, 2011 to December, 2012 in China Rehabilitation Research Center were randomly divided into control group (n=15), neuromuscular electrical stimulation (NMES) group (n=15) and ETNS group (n=15). 3 groups were given routine medication and rehabilitation treatment. They were assessed with the largest surface electromyography (sEMG), Simple Test for Evaluating Hand Function (STEF), and modified Barthel Index before and after treatment. Results After treatment, the range of sEMG of extension carpi radialis and STEF improved in three groups (P<0.05). NMES group and ETNS group were better than the control group (P<0.05), and ETNS group was better than NMES group (P<0.05). The scores of modified Barthel Index rose (P<0.05), NMES group and ETNS group were better than the control group (P<0.05). Conclusion Both NMES and ETNS can improve the motor recovery of upper limbs after stroke, and ETNS is more effective.

Background The integration of segregated pathways from the two eyes first appears in V1 neurons,where it not only plays a critical role in the generation of a three-dimensional visual representation.Abnormal visual experiences in critical period usually lead to amblyopia and binocular integration defects.Objective Present study was to investigate how neurons of kitten coordinate their activity patterns in response to synchronous dichoptic stimulus inputs in striate cortex.Methods Spike rate and local field potential(LFP) gamma band(20-90Hz) power of three kitten(1-1.2Kg,8-10 weeks old) to monocular and synchronous dichoptic presented gratings were assessed for 28 binocular neurons in V1 of kitten by in vivo extracellular record method under anaesthesia and paralysis.Ocular dominance index(ODI) and binocular integration index(BII) were assessed and the correlation between these two indexes were analyzed.Results In 28 cells with binocular characteristic,the absolute value of spike-ODI was significant larger than that of LFP-ODI(t=2.606,P=0.021).A positive linear correlation between the ocular preferences of spike and LFP was found(R2=0.513,F=27.423,P=0.003).In dichotic trails,binocular facilitation with BII for spike was 2.348±0.996,showing a significant reduce in comparison with BII for LFP(3.678±1.974)(t=2.671,P=0.019).Binocular integration index for two signals were greater when monocular responses of both eyes were similar(P=0.035 and P=0.124,respectively).Conclusion Both spike rate and gamma band power of LFP exhibited binocular facilitation to synchronous presented dichotic stimuli with significant facilitation induced by balanced monocular responses.Spiking activity and LFP reflect neural activities of different spatial scales and source components.

Objective To establish a real-time quantitative PCR ( qPCR) method for detection of Streptobacillus moniliformis, which can be used to rapidly detect this pathogen in laboratory animals .Method According to the S. moniliformis sequences published in NCBI , we designed specific primers and MGB probe .The specificity, sensitivity and stability of this method were evaluated using 24 standard reference strains .Total of 823 respiratory specimens of animals including mice, rats, guinea pigs, hamsters, rabbits, Mongolian gerbils and tree shrews , were detected by this established Taqman MGB qPCR method .Results We had successfully established the S.moniliformis Taqman MGB qPCR method . S.moniliformis was not detected in the samples of mice , rats, guinea pigs, hamsters and rabbits.The positive rate of S. moniliformis was 1.5% ( 1/65 ) and 61.7% ( 37/60 ) in conventional Mongolian Gerbils and tree shrews , respectively . Conclusions Our developed qPCR method can be used to effectively detect S.moniliformis in laboratory animals .Moreover , its accuracy and sensitivity are better than the national standard method .This study laid the foundations for optimizing the quality inspection system of laboratory animals .

Objective To establish an effective PCR assay for detection of Bartonella, and application of this assay in tree shrew .Methods Sequence of Bartonella was obtained from NCBI Genbank .Three pairs of primers were designed based on this sequence .One pair of primers was determined through amplifying the major strains in China .Sixty tree shrew blood samples were tested with this PCR assay .The positive amplified fragments were sequenced to verify the reliability of this method .Results A PCR method for detection of Bartonella is successfully established , with a high specificity and the sensitivity was of 2.0 ×10 -5 μg/mL.Among the tested 60 blood samples , 15 positive cases were detected.Sequencing of the samples confirmed a 25%infection rate of Bartonella in the tree shrews, well consistent with the amplification results , and verified the applicability of this detection method .Conclusion The establishment of this method provides the basis for detection of Bartonella in tree shrew.

Objective To explore the incidence and clinical characteristics of Han and Uygur pa- tients with ulcerative colitis(UC)in xinjiang.Methods Six hundred and ninty nine patients with UC from 2000 to 2006 were retrospectively reviewed.Patients were divided into 2 groups according to ethnic back- ground,including 522 Han patients and 177 Uygur patients.General data,age,clinical manifestation,le- sion range,lesion degree and complications were recorded in the 2 groups respectively.Results The detec- tion rate of UC by endoscopy in Urgur group was higher than that in Han group(P