2.Develonment and application of TaqMan MGB probe real-time fluorescence quantitative PCR for rapid detection of Helicobacter hepaticus
Zhengqin GAO ; Jin XING ; Yufang FENG ; Bingfei YUE ; Zhengming HE
Chinese Journal of Microbiology and Immunology 2011;31(9):833-838
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.
3.Influence of Jiangzhipailuan decoction on the related factors of lipid metabolism in patients with polycystic ovary syndrome
Yufang WANG ; Dongyun KE ; Peizhu FENG ; Jiangqiong LUO ; Li TAN
Chinese Journal of Primary Medicine and Pharmacy 2011;18(1):18-20
Objective To explore the effect of Jiangzhipailuan decoction in regulating PPARα( belong to the nuclear receptor family of ligand-activated transcription factors ), PGC- 1α ( peroxime proliferator activated releptour)and SREBP-1c( belong to the baichelix-loop-helix-leucine zipper class of transcription factors), SCAP( SREBP cleavage activating protein) related to lipid metabolism in the treatment of polycystic ovarian syndrome. Methods 84 patients suffered polycystic ovary syndrome were randomly divided into 4 groups: one received traditional Chinese medicine treatment, one western medicine treatment, one combination therapy and one as the control group. Traditional Chinese medicine group was treated with Jiangzhipailuan decoction treatment for 3 months, western medicine group was treated with up to Diane-35 ( ethinyl cyproterone tablets) for three cycles, while the combination therapy group was treated with traditional Chinese medicine ,western medicine as well as combined treatment for 3 courses. Results In the combination therapy group the PPARα, PGC-1α and decreased SREBP-1c, SCAP copy number was significantly improved ,and body mass index was significantly lowered. The total improvement rate of menstrual in three groups were 71.42% ,75.00% ,92.86% respectively. Conclusion Jiangzhipailuan decoction played a prominent role in regulating PPARα,PGC-1α and SREBP-1c,SCAP related to lipid metabolism in the treatment of polycystic ovarian syndrome.
4.Development and application of a multiplex PCR assay for pathogenic dermatophytes in laboratory animals
Jin XING ; Yufang FENG ; Bingfei YUE ; Zhengming HE
Chinese Journal of Comparative Medicine 2015;(12):65-70
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.
5.Phenotypic analysis of Pasteurella pneumotropica in laboratory animals in Beijing area
Jin XING ; Yufang FENG ; Bingfei YUE ; Zhengming HE
Chinese Journal of Comparative Medicine 2014;(6):54-57
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 .
6.Spindle cell carcinoma of the breast:a clinicopathologic analysis of three cases
Zhongyan GUAN ; Yufang FENG ; Zhenkang GAN ; Liang SHI ; Yingyong HOU
Chinese Journal of Clinical and Experimental Pathology 2015;(9):996-1000
Purpose To analyze the clinical pathological characteristics and pathological diagnosis and prognosis of spindle cell carci-noma of the breast. Methods Three cases of spindle cell carcinoma of the breast were studied by morphological and immunohisto-chemical EnVision techniques. Results The females were 48, 63 and 71 years old. The tumors located in the right breast with 4. 0 cm × 3. 0 cm × 3. 0 cm, 3. 0 cm × 2. 0 cm × 2. 0 cm and 3. 5 cm × 2. 8 cm × 2. 3 cm in size and showed cystic lesion. The neoplasm was composed of bland spindle cells and mimicking fibromatosis. Immunohistochemical staining showed that spindle cells were positive for CK(AE1/AE3), CK(34βE12), CK14, CK5/6, p63 and vimentin, negative for ER, PR and c-erbB-2. Ki-67 was positive in 20%, 25% and 20% of the cells. Conclusion Spindle cell carcinoma of the breast is a rare subtype of the metaplastic carcinoma which tend to show cystic changes. It is important to make a definite diagnosis which combine histopathologic features and immunophe-notyping.
7.A comparative study of the drug negotiation mechanism in typical regions at home and abroad
Rui NIU ; Ying LIU ; Yufang XIANG ; Bianling FENG
Chinese Journal of Health Policy 2017;10(6):25-32
With this retrospective study, we analyzed the negotiation subject, drug type, drug access mechanism, negotiation link and negotiation result of drug negotiation in typical regions at home and abroad.In foreign countries, drug negotiation mechanism has been implemented for many years, and many countries have established a relatively perfect negotiation system.We selected 8 typical countries and regions for which we statistically analyzed the negotiation subject, drug type, drug access mechanism, negotiation link and negotiation result of drug negotiation and these include the United States, Australia, Canada, France, Germany, Italy, South Korea and Taiwan.However, in recent years some Chinese medical insurance department carried out pilot works on drug negotiations, and they got some successful results of the implementation of practical experience.Eight typical Chinese regions were also selected for statistical analysis, and these include Zhejiang, Hunan, Jiangsu, Jiangxi, Chengdu and Qingdao.From the analysis of the comparison of drug negotiation mechanism in domestic and foreign typical regional, we found that foreign regional drug negotiation mechanism is more mature and perfect, while in the domestic areas the mechanism is still poor at a certain extent as compared to foreign countries.We should learn from the successful experience of foreign countries and also establish and improve the negotiation mechanism that is suitable for China''s national conditions.
8.Investigation of Leptospira infection in three new experimental animals by PCR methods
Yufang FENG ; Jin XING ; Wei GONG ; Bingfei YUE ; Zhengming HE
Chinese Journal of Comparative Medicine 2014;(8):31-35,66
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 .
9.Relation between transvaginal color Doppler sonographic findings and the medical conservative treatment results of tubal pregnancy
Qin XUE ; Zhimin ZHANG ; Dazhong ZOU ; Yufang XU ; Hong YU ; Jie TAN ; Feng XU
Chinese Journal of Ultrasonography 2009;18(1):59-61
Objective To evaluate the value of transvaginal color Doppler sonography(TVCD)in the conservative treatment of early tubal pregnancy(TP).Methods Fifty cases of early TP were examined by TVCD before medical treatment,including the size of TP mass,blood flows graded according to Alder,hemodynamics parameters.All data were ananlized and compared with therapeutic results.Results Fortyfive cases were treated successfully(45/50),and 5 failed.According to TVCD,TP masses flow were graded from O to Ⅲ.In the successful group,4 cases were graded 0,21 Ⅰ,16 Ⅱ and 4Ⅲ,blood flow signals were measured in 41 cases,the mean velocity was(5.452±4.327)cm/s,PI(1.597±0.696),RI 0.680±0.107.In the failure group,all TP masses flow were graded Ⅲ,the mean velocity was(16.774±9.855)cm/s,PI 0.95 1±0.193,RI 0.567±0.034.Conclusions In the medical treatment of early TP,TVCD findings associated with the treatment outcome closely,it plays an important role in assessing conservative treatment of early TP.
10.Results and analysis of the proficiency of laboratories in detection of Salmonella in laboratory animals
Jin XING ; Yufang FENG ; Hong WANG ; Ji WANG ; Bingfei YUE ; Zhengming HE
Acta Laboratorium Animalis Scientia Sinica 2016;24(2):191-194
Objective To understand the Salmonella detectability in the laboratory animal testing laboratories, im-prove the level of detection for the quality of laboratory animals, by means of laboratory animals Salmonella proficiency tes-ting program.Method According to the proficiency testing program approved by CNAS, freeze-dried animal stool samples containing Salmonella bacteria and interference bacteria were prepared, and through stability and homogeneity tests quali-fied as proficiency testing samples.The randomly numbered samples were issued to the participating units by cold-chain transportation, and attached work instructions.The original reports and copies of the tests should be submitted on time.The sample results consistent with the results of the pretested results were considered as satisfactory, and the results inconsistent or fails to submit were judged as unsatisfactory.Results A total of 30 laboratories from 20 provinces and cities nationwide participated in this proficiency testing programs for Salmonella, including 28 ( 93.3%) laboratories with satisfactory re-sults, and two laboratories unsatisfactory ( 6.7%) .29 laboratories used separate culture methods, and two laboratories used PCR method.Conclusions The laboratory animal quality inspection agencies have good detection ability for Salmo-nella.The implementation of the capacity verification plan can well reflect the detection level of laboratories.