Animals ; Cell Line, Tumor ; Female ; Humans ; Mammary Neoplasms, Experimental ; diagnosis ; metabolism ; pathology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Organometallic Compounds ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Radioisotopes ; Radiopharmaceuticals ; pharmacology ; Random Allocation ; Rhenium ; Tumor Burden ; drug effects ; bcl-2-Associated X Protein ; metabolism

Adolescent ; Bone Marrow Transplantation ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Thalassemia ; surgery ; Transplantation Chimera

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.

Autoimmune Lymphoproliferative Syndrome ; diagnosis ; genetics ; immunology ; therapy ; Biomarkers ; blood ; Child, Preschool ; Humans ; Male ; Mutation ; Prednisolone ; administration & dosage ; therapeutic use ; Ultrasonography, Doppler, Color ; fas Receptor ; genetics

Objective:To evaluate the accuracy of the application of color Doppler sonography (CDS) and computer tomography angiography (CTA) in preoperative perforator identification and flap design and provide theoretical support for the restoration of oral maxillofacial defect with free superficial circumflex iliac artery perforator flap (SCIAPF). Methods: (1) Preoperative CDS and CTA techniques were performed to map the SCIA perforators of 29 adult patients diagnosed with malignant tumor in the oral maxillofacial head and neck regions. These patients were scheduled for concurrent reconstruction surgery. (2) A diagnostic test was designed to com-pare the CDS and CTA techniques. Results:(1) A total of 18 patients underwent flap preparation. SCIA was not found in one of the pa-tients during surgery, but was observed intra-operatively in the other 17 patients. The average SCIA diameter was 0.69 ± 0.20 mm. (2) The diagnostic test showed a CDS sensitivity of 75.0%, a CDS specificity of 82.4%, and an area under the ROC curve of 0.79. The CTA sensitivity was 75.0%, the specificity was 94.2%, and the area under the ROC curve was 0.85. The diameters measured by CDS and CTA were compared with the diameter measured intra-operatively. Significant differences were observed among the three diame-ters (P<0.05). The average diameter measured by CDS was 0.84 ± 0.14 mm. The average diameter measured by CTA was 1.01 ± 0.19 mm. Conclusion:CDS and CTA are relatively reliable technologies for preoperative detection of perforator vessel. The use of CDS and CTA technology mapping for SCIAPF can provide accurate information about the perforator, including the position of the perforator and the relationship between the peripheral tissues and the caliber of the vessel.

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 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 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 .

ObjectiveTo observe the effect of intravesical instillation of Brucea Javanica oil emulsion on recurrence of bladder cancer after transurethral resection of bladder tumor (TUR-Bt) operation.Methods187 patients with superficial bladder carcinoma after TUR-Bt operation were randomly divided into the group A (85 cases) and group B (102 cases). Patients of the group A were treated with instillation of Brucea Javanica oil emulsion; those of the group B were treated with mitomycin. A three-years following up was performed to observe the recurrence and side effects.ResultsAfter a 3-years following up, the recurrence rate of group A was 12.94%, lower than that of group B (34.31%). The side effects were seldom seen in the group A.ConclusionThe effect of intravesical instillation of Brucea Javanica oil emulsion to prevent the recurrence of bladder cancer after TUR-Bt operation is favorable.

Objective To investigate the effects of fragile X mental retardation protein（FMRP）on the development and migration of cerebellar neurons in mouse model.Methods Plasmids containing FMRPmutant-EGFP or EGFP were established and transfected into the lateral ventricle of the embryo mouse.Fragile X syndrome（FXS）genotype of the mouse model was identified.Nissl staining and immunofluorescence staining were conducted to assess the changes in neuron development and migration.Results In the experimental group,Nissl staining showed that the deep cerebellar neuclei contracted and divided by white matter,and the non-polarized Purkinje cells retained in internal granular layer;while immunofluorescence staining showed that Tbr2-positive unipolar brush cells changed the migration pathway and accumulated in the ventricular zone.Conclusion Cerebellar neurons showed abnormal formation and migration with the absence of FMRP.