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.

Objective: To compare and analyze the antimicrobial efficacy of three different mechanical preparation techniques in single infected root canals. Methods; Forty-five single root canals with chronic periapical periodontitis were selected. The specimens were divided into three groups randomly, 15 root canals per group. Croup A: preparation with stainless steel K-files (step-back technique), Croup B: preparation with HERO 642 NiTi rotary files (crown-down technique) and Group C: preparation with Mtwo NiTi rotary files ( modified crown-down technique). The sterile normal saline was used as irrigation. Samples were taken before and after canal preparation. The difference of CFU was calculated as well as the bacterial species. Results; All groups were effective to reduce bacteria within the infected root canals greatly(P<0.01). Croup A and Group C were statistically better than Group B(P<0.05). Group A was more effective than Group C but there was no statistically difference between them(P>0.05). Conclusion; Mechanical preparation can greatly reduce the intracanal bacteria, but can not obtain bacteria-free canals. The mechanical preparation must be aided by chemical irrigation to improve the success of root canal therapy.

[Objective]To evaluate whether transplanted marrow mesenchymal stem cells interfered in TGF-?1 can differentiate to nucleus pulposus cells and increase the amount of proteoglycan and collagenase Ⅱ content in intervertebral discs.[Method]We used an in vivo model to investigate the feasibility of marrow mesenchymal stem cells that cultured in vitro and interfered in TGF-?1 delivery,retention,and survival in the degeneratived disc space.In 2,4,6,8 weeks we used immunohistochemical staining to determine the change of collagenase Ⅱ content;spectrophotometry to determine the change of amount of proteoglycan with Phlorglucinol;the experiment date were analyzed by SPSS 11.5 soft ware.[Result]We found MSCs could maintain viability and proliferate within the rabbit inter vertebral disc.The amount of proteoglycan and collagen Type Ⅱ content of the intervertebral in matrix synthesis in the experiment group was increased in 8 weeks.We found no changes in the modle group.[Conclusion]Our data suggest that transplanted marrow mesenchymal stem cells in vivo can survive and increase proteoglycan and collagen Type Ⅱ amount interfered in TGF-?1 in some periods,which support its potential use as a treatment of intervertebral disc degeneration.

[Objective]To establish an in vitro two-dimensional culture model of degenerated nucleus pulposus and detect samples' cell cycle by flow cytometry to study why nucleus pulposus cells don't grow well after passage. [Method]Nucleus pulposus tissues taken from protruded discs of 24 patients were treated by Trypsin and collagenase after surgical procedures,and then the cells were cultured in DMEM/F12 medium.Cell morphology was observed by an invert microscope and cell cycle of the primary and P2 cells were detected by flow cytometry after proliferation in monolayer culture.[Result]1.Primary culture cells of nucleus pulposus grew well in the medium,and 90% cells adhere to layer after about 7d.2.The rate of apoptosis of NP: primary(38.10?11.7)%,P2(44.74?17.6)%.The rate of S period : primary(7.88?2.1%),P2(2.76?0.7)%.[Conclusion]When going down to posterity the cells' apoptosis rate grows while S period cells decrease.

Objective To investigate the operative technique and treatment effect for the cerebellum he-mangioblastoma. Methods The clinical data of 18 patients with cerebellum hemangioblastoma who underwent sur-gery were analyzed retrospectively. The clinical data included imaging data,the operative approach and microsurgery method. Results In the 18 cases, 15 undertook complete remove of tumor,and the other 3 cases experienced partial remove. 16 patients were followed up for 1 to 3 years,and 2 cases experienced recurrence. No death occurred. Con-clusion The measures including sufficient preoperative imaging and gentle manipulation during the operation can significantly increase the rate of total removal of the tumors, while radioneurosurgery or γ treatment is applicable after operation.

Objective To investigate mesenchymal stem cells genetically modified by nerve growth factor to repair acute spinal cord injury in rats. Methods Fifty six Wistar rats of inbred strain were randomly divided into sham operation group cell transplantation group and simple injury group. The spinal cord injury model was prepared according to the modified Allen's method. NGFβ (hNGFβ) and GFP genes were transfected into MSCs by replication-deficient recombinant adenovirus vector (Ad-hNGFβ) and replication-deficient recombinant retrovirus vector (Rt-GFP) respectively. GFP positive MSCs were transplanted into intradural space of injured spinal cord at 7 days after spina coral injury. Spinal cord was dissected at 24 h, 1 and 2 weeks after transplantation. To observe the expression of GFAP and nestin and the distribution of MSCs after transplantation following the spina corol injury. Results MSCs migrated to the injured parenchyma In transplantation group, the expression of GFAP and NGF protein was higher than in the control group (P<0.05), the BBB score in transplantation group was higher than that in the control group (P<0.05). Conclusion The MSCs transplantation repaired the injured spinal cord to some extent.

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.

Objective To understand the epidemic situation and characteristics of malaria in Yunnan Province,so as to pro?vide the reference for malaria elimination. Methods The data of malaria reported in the information system were collected and analyzed in Yunnan Province from 2011 to 2013. Results From 2011 to 2013,totally 2 256 malaria cases were found in Yun?nan Province,with a morbidity of 0.162 8 per million and three of them were death cases. The local cases mainly distributed along the boundary and accounted for 29.48%,while the imported cases mainly came from Myanmar and accounted for 70.52%. The number of endemic counties with local malaria cases decreased from 37 to 10 during the three years. The number of import?ed cases reached the peak in May and the local cases in June. The patients were mainly aged from 20 to 49 years old(accounted for 70.58%),and 85.24% of the cases were peasants and laborers. Totally 86.66% of cases were laboratory confirmed cases, and 13.14% were clinically diagnosed. The proportions of cases reported by hospitals,health service centers and CDCs were 33.02%,37.06% and 29.92%,respectively. Conclusions The prevalence of malaria in Yunnan Province decreased from 2011 to 2013. The work of malaria cases double?checked by province?level CDCs is effective. However,the awareness and accurately diagnostic capability of clinical doctors still should be strengthened.

Objective:To analyze the causes and risk factors for failure of internal fixation with proximal femoral nail antirotation (PFNA) in the treatment of femoral intertrochanteric fractures.Methods:A retrospective analysis was conducted of the 568 patients with femoral intertrochanteric fracture who had been treated with PFNA fixation at Department of Orthopaedic Surgery, The Fifth Central Hospital of Tianjin from March 2013 to March 2018. They were 348 males and 220 females, aged from 44 to 93 years (average, 74.6 years). According to the fracture stability classification, the patients were divided into a stable group of 424 cases and an unstable group of 144 cases. According to the AO classification, the stable group had type 31-A1 and type 31-A2.1 while the unstable group type 31-A2.2, type 31-A2.3 and type 31-A3. The 2 groups were compared in terms of reduction quality, rate of internal fixation failure, and function of the affected hip. Single factor and multi-factor binary logistic regression analyses were conducted to determine the risk factors responsible for failure of PFNA fixation of femoral intertrochanteric fracture.Results:There were no significant differences in the preoperative general data between the 2 groups, showing comparability between groups ( P> 0.05). Internal fixation failure occurred in 19 cases, which was caused by spiral blade withdrawal in 13 cases, femoral neck shortening in 17 cases, hip varus in 14 cases, and spiral blade cut-out in 14 cases. The failure rate for the stable group was 1.2% (5/424), significantly lower than that for the unstable group [9.7%,(14/144)] ( P<0.05). The Harris hip score at the last follow-up for the stable group [98(95,100)] was significantly higher than that for the unstable group [84 (82, 87)] ( P<0.05). There was no significant difference in reduction quality between the 2 groups ( P>0.05). The multivariate analysis showed that osteoporosis ( OR=7.283, 95% CI: 1.626 to 32.623, P=0.009) and unstable fracture ( OR=11.607, 95% CI: 4.039 to 33.355, P<0.001) were risk factors responsible for the failure of PFNA fixation of femoral intertrochanteric fracture. Conclusions:PFNA fixation for unstable intertrochanteric fracture can lead to a high failure rate. It forms a lever like structure so that the main stress is shifted to the internal fixation. Its lever fulcrum is located at the angle of intramedullary fixation so that a long arm forms at the load-bearing side, leading to a high failure rate. The weight-free time should be longer for patients with osteoporosis and unstable fracture after operation.