Objectives To establish a sensitive and special method for the detection of Ureaplasma urealyticum(Uu) using PCR microplate hybridization (PCR MPH). Methods A primer of ureasea gene was labeled by biotin. The amplification product was captured on streptavidin coated microplates, then products were quantified by hybridization with a digoxigenin labeled internal oligonucleotide probe. After revelation with an anti digoxigenin alkaline phosphatase coupled antibody(anti DIG AP), the amount was determined by optical reading. At the same time, PCR MPH was compared with Bio Merieux Mycosplasma IST. Results A method of PCR MPH for detecting Uu DNA was established. The morbidity among three groups for detecting 158 clinical samples was analysed. 65 were detected by PCR MPH and 56 by culture.Conclusion The results showed that this assay is rapid, sensitive, specific, and accurate, and is of value in clinical therapy.

50.0%).ESBLs isolates reached up 63.5%.CONCLUSIONS E.coli has a high detection rate and serious drug resistance in urinary infections.Conventional ESBLs tests can help to appropriate use antibiotics and lower the occurrence of drug resistance.Clinical doctors are advised to restrict the usage of antibiotics in order to lower the occurrence of drug resistance strains,as well as to prevent the spread of ESBLs.

Objective To investigate infection statue and antibiotic susceptibility of Mycoplasma urealyticum and Mycoplasma hominis in the genitourinary tract in our area, so as to instruct the rational use of antibiotics. Methods Genitourinary secretions were collected with swabs. They were cultured with the diagnostic kit of Mycoplasma (Biomerieux Company) to detect M. urealyticum and M. hominis. Meanwhile the susceptibility of Mycoplasma against 9 antimicrobial agents was tested with the same kit. According to the manual of the kit,the results were read. The data were statistically analyzed. Results A total of 2 410 samples were collected, and the positive rate was 58. 1%. Among 1410 positive cases of Mycoplasma, 901 cases were M. urealyticum (37.4%), 85 were M. hominis (3.5%), and 415 were M. urealyticum combined with M. hominis (17.2%).The susceptibility rate of M. urealyticum to josamycin, pristinamycin, ciprofloxacin was 98. 8%, 98. 8% and 6. 4% respectively, while the susceptibility rate of M. urealyticum combined with M. horninis was 86.9%,86. 8% and 2.6% respectively. Conclusion M. urealyticum is the major cause of Mycoplasma infection in genitourinary system. Josamycin and pfistinamycin are more effective than other antimicrobial agents to treat Mycoplasma irffection. Ciprofloxacin is more resistant than other antimicrobial agents. Sensitive antibiotics should be selected based on the results of bacterial culture and drug sensitivity tests so as to raise the clinical curative effects.

To explore the role of connexin43 (Cx43) in gap junctional intercellular communication (GJIC) and propagated sensation along meridians, the expression of Cx43 in the rat epithelial cells and fibroblasts was studied both in vitro and in vivo. With the in vitro study, the rat epithelial cells and fibroblasts were cultured together, and the localization of Cx43 was detected by immunohistochemistry and indirect immunofluorescent cytochemistry and under confocal microscopy. And the expression of Cx43 on the surface of the cells was examined by flow cytometry. With the in vivo examination, 20 SD rats were randomized into control group (n = 10) and electrical acupuncture group (EA group, n = 10). EA ( 0.5-1. 5 V, 4-16 Hz , 30 min) was applied to "Zusanli" acupoint for 30 min at rat's hind paw, the localization of Cx43 was immunohistochemically detected. The immunohistochemical staining and indirect immunfluorescent cytochemistry showed that Cx43 was localized on the surface of the cells and in the cytoplasm. The relative expression level of Cx43 on the cellular membrane surfaces of the rat epithelial cells and fibroblasts, as determined by FACS, were 13.91% and 29.53% respectively. Our studied suggested that Cx43 might be involved in GJIC and propagated sensation along meridians.
Cells, Cultured ; Connexin 43/*biosynthesis ; Connexin 43/genetics ; Epithelial Cells/cytology ; Epithelial Cells/*metabolism ; Fibroblasts/cytology ; Fibroblasts/*metabolism ; Flow Cytometry ; Gap Junctions ; Meridians ; Microscopy, Confocal ; Random Allocation ; Rats, Sprague-Dawley

Objective To evaluate the advantages of cytomegalovirus (CMV) real-time PCR, to monitor CMV infection in a population of transplantation recipients in comparison with the qualitative PCR method.Methods 150 plasma samples and leukocytes samples collected from 59 bone marrow transplant recipients and 9 liver transplant recipients were tested by qualitative CMV PCR assay in parallel; Real-time CMV PCR using Roche Light Cycler was performed with 150 plasma samples and 54 control plasma samples.Results Using real-time PCR as the reference standard, the sensitivity and specificity of the qualitative CMV PCR assay with leukocytes samples were 62.5% and 95.5%, with plasma samples were 57.5% and 99.1% respectively. The recipients with the level of CMV DNA over 5?10~3 copies/ml by real-time PCR had higher percentage of developing CMV disease than those below 10~3 copies/ml.Conclusion The quantitative detection of CMV DNA by real-time PCR with plasma is a rapid, specific and sensitive method to monitor CMV infection in patients after transplantation and to guide antivirus therapy.

Objective To explore human cytomegalovirus UL97 mutations related to ganciclovir resistance in hematopoietic stem cell transplant (HSCT) recipients.Methods A total of 43 patients,including 24 males and 19 females,with an average age of 21 years old,who had HCMV DNAemia for more than two weeks after HSCT between 2008 and 2010 in Peking University People's Hospital,were included in this prospective study.UL97 GCV resistance mutations were investigated in 49 plasma specimens collected from those patients.GCV resistance mutations such as UL97 M460V/I,H520Q,A591V,A594V,L595S/F,and C603W,were analyzed by modified PCR-RFLP methods.UL97 mutations related to GCV resistance were assayed by the method of PCR-direct sequencing (PCR-DS).An amplified refractory mutation system real-time PCR (ARMS RT-PCR) was developed for the detection of UL97 A594V mutation.Results Eight known UL97 ganciclovir resistance mutations were not detected by PCR-RFLP and PCR-DS.Four new UL97 mutations like UL97 R494P,T502A,N558D,and G561S,were detected by PCR-DS.The ARMS RT-PCR for detecting of UL97 A594V was established successfully.The lower limit of detection of the method was at least 7.5 × 102 copies/ml combined with the use of nucleic acid extraction reagent.UL97 A594V resistance mutation was identified by the method of ARMS RT-PCR in two HSCT recipients.The rate of UL97 A594V mutation was 4.7％ (2/43) in HSCT recipients.Conclusion The ARMS RT-PCR assay represented a sensitive method for the identification of UL97 A594V mutation.

r diagnosing the disease to combine pathology, immunohistochemistry and SYT-SSX gene detection.

Objective To study the clinical characteristics of tuberculosis in oral and maxillofacial region,and to provide reference in early diagnosis and early treatment. Methods The clinical data of 132 patients were analyzed retrospectively. ResultsIn all cases, male to female ratio of 1∶1.69, the average age of 38.7 years, the group of 15 ～40year-old had higher incidence. Al1 parts of oral and maxillofacial region could be involved. In all 132 cases,107 cases were treated by surgery and drugs,25 cases by drugs only. Conclusion The local sign of tuberculosis in oral and maxillofacial region was atypical ,so it was likely to misdiagnosis. Definitive diagnosis relied on pathological examination. Operations played an important role in the cases of unifocal lymph nodes tuberculosis,oral mucosal tuberculosis, multifocal concentrated lymph nodes tuberculosis and salivary tuberculosis, while the therapy of multifocal sporadic lymph nodes tuberculosis and osseous tuberculosis should be relied on the medicine.

Objective To investigate CT and MRI features of dermatofibrosarcoma protuberans (DFSP).Methods Totally 16 patients with DFSP confirmed by pathology were enrolled.Tumor morphology,CT and MRI imaging appearance (11 cases underwent plain and enhanced CT,5 cases underwent plain and enhanced MRI) were analyzed retrospectively.Results DFSP usually occurred in the skin of truck,head and neck,protruding from the skin surface in different extent.Some lesions even suspended out of the skin.The lesions were divided into nodular type (n=12) and diffuse type (n=4) according to their morphological appearance.The tumors usually demonstrated as iso-density or slightly low density solid mass compared to muscle on CT.On MRI,it usually demonstrated as low signal on T1WI and high signal on T2WI.Tumor blood supply was rich,and it usually showed progressively moderate to strong enhancement.The signs within DFSP include hanging sign (n=2),skin tail sign (n＝6),fascia tail sign (n=l),fat tail sign (n=4).Conclusion DFSP can be characterized by nodular or diffuse lesions,the manifestations of different form are slightly different,but still have a certain characteristic.

To explore the role of connexin43 (Cx43) in gap junctional intercellular communication (GJIC) and propagated sensation along meridians, the expression of Cx43 in the rat epithelial cells and fibroblasts was studied both in vitro and in vivo. With the in vitro study, the rat epithelial cells and fibroblasts were cultured together, and the localization of Cx43 was detected by immunohistochemistry and indirect immunofluorescent cytochemistry and under confocal microscopy . And the expression of Cx43 on the surface of the cells was examined by flow cytometry. With the in vivo examination, 20 SD rats were randomized into control group (n = 10) and electrical acupuncture group (EAgroup, n=10). EA ( 0.5-1.5 V, 4-16 Hz , 30 min) was applied to"Zusanli"acupoint for 30 min at rat's hind paw, the localization of Cx43 was immunohistochemically detected.The immunohistochemical staining and indirect immunfluorescent cytochemistry showed that Cx43was localized on the surface of the cells and in the cytoplasm. The relative expression level of Cx43on the cellular membrane surfaces of the rat epithelial cells and fibroblasts, as determined by FACS, were 13.91 % and 29.53 % respectively. Our studied suggested that Cx43 might be involved in GJIC and propagated sensation along meridians.