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 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 observe therapeutic effects of early scab-cutting combined with axis-like abdo-men skin flap implanting in patients with wrist deep electrical burns.Methods Retrospective study was employed to analyze 70 patients received axis -like abdomen skin flap implanting for wrist deep electrical burns.All the cases were separated with early scab-cutting group(35 cases)and delayed scab-cutting group(35 cases),according to types of skin flap implanting.The therapeutic effect for the two groups were evaluated with rate of infection,duration of healing,living rate of skin flap,grade of skin burning,and rate of amputate.Results In the early scab -cutting group,the rate of infection was 11%,the duration of healing was(24.0 ±6.0)days,the rate of amputate was 3%,the living rate of skin flap was 97.0%,the wound healing of class A was 94.3%.In the delayed scab-cutting group,the data were respectively 31%,(35.0 ±7.4)days,17.0%,31% and 28.6%.Compared with the data of the two groups,the difference had statistical significance(the chi-square value or T value were respectively 4.151,6.831,3. 968,31.895,3.968.All the P values was less than 0.05).Conclusion Early scab-cutting combined with varied sorts of skin flap implanting shown great benefits for treatment of wounded and facilitate rehabilitation in function of wrist,which suggest decrease rate of complications and minimized hospitalized duration.

Objective To observe the clinical effect of the anterolateral thigh flap in repairing defect caused by resection of malignant melanoma in the heel region.Methods 8 patients with heel malignant melanoma were trea-ted by surgery.The malignant melanoma was further diagnosed by histopathological examination.Their heel defects caused by large resection of malignant melanoma were treated by anterolateral thigh flap.Results All skin flaps sur-vived and all the patients could walk normally 1 month after operation.The recovery of the function and sense was well 6 months after operation.Compared with the another well heel,the flaps had normal texture,color and appearance of heel.8 patients had complete follow -up,ranged form 6 months to 3 years.All cases survived with neither local recur-rence nor distant metastasis.Conclusion The anterolateral thigh flap can achieve good clinical effect in treating heel defect caused by the resection of malignant melamoma.And especially apply in large skin and tissue defect of heel area.The flap can weight,wear -resisting and reduce overheating and over cold damage.The range of resection can not be restricted by local skin flap.The clinical effect is satisfied.

Objective: To compare the efficacy of casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) and Duraphat varnish on preventing radiation caries and dentin hypersensitivity (DH) in irradiated head and neck cancer pa-tients. Methods:A total of 112 patients with head and neck cancer who will undergo radiotherapy were selected and randomly divided into the CPP-ACP group (experimental group) and the Duraphat varnish group (control group). A day prior to the scheduled radiothera-py, CPP-ACP containing paste had been applied once a day for 5 weeks to the teeth surface of the experimental group. Duraphat varnish was applied with the same frequency in the control group. The decay missing filling tooth (DMFT), decay missing filling surface (DMFS), and DH of two groups before radiotherapy and 12 months after radiotherapy were observed and statistically analyzed. Re-sults:Before radiotherapy, DMFT, DMFS, and DH showed no significant differences between the groups (P>0.05). However, signifi-cant increase in DMFT, DMFS, and DH was noted in the two groups 12 months after the radiotherapy (P<0.05). A higher increase was observed in the DMFT, DMFS, and DH of the control group compared with those in the experimental group (P<0.05). Conclusion:CPP-ACP effectively reduced radiation caries and sensitivity in irradiated head and neck cancer patients and deservesclinical applica-tion.

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 explore the predictive value of serum CEA and cytokeratin-19 fragments (CYFRA21-1)prior treatment for the epidermal growth factor receptor (EGFR) mutation and efficacy of tyrosine kinase inhibitors ( TKI ) in patients with non-small cell lung cancer.Methods The study was a clinical research.Totally 101 matched tissue and plasma samples were collected from Peking University People′s Hospital from 2012 to 2013.All clinical specimens were analyzed for EGFR mutations in exons of 18, 19, 20 and 21 by ADx-ARMS and direct sequencing, and the serum levels of CEA and CYFRA21-1 were analyzed by ECLI.The correlation between EGFR mutant status and efficacy of EGFR-TKI and clinicopathological parameters were analyzed by χ2 test, Log-rank text and Cox proportional hazards regression model.Results The mutation rate was 60.4%(61/101) by ADx-ARMS and 33.7%(34/101) by direct sequencing.Mutations were more frequently observed in the higher serum CEA level patients(≥5μg/L,78.8%).However, the rates of EGFR mutations of different CEA levels were similar.Among the patients receiving TKI therapy, the efficacy of EGFR-TKI was closely related to serum CYFRA21-1 level prior treatment and EGFR mutation (χ2 =8.903, P =0.003; χ2 =28.590, P <0.001 ).And serum CYFRA21-1 level prior treatment and EGFR mutation were independent factors for EGFR-TKI treatment affecting PFS (RR=0.298, P<0.001;RR=0.086, P<0.001).Conclusion The mutation rate of EGFR was significantly related with the expression level of CEA prior treatment, and serum CEA and CYFRA21-1 levels prior treatment could be potential predictors of EGFR-TKI efficacy.

Objective To observe and analyze the clinical curative effect of the platelet rich plasma (PRP) in treatment of chronic difficult to heal the wound.Methods We selected 60 patients with chronic difficult healing wounds and they were divide into two groups at random,30 patients of A group,B group of 30 patients.All patients were given routine debridement,drainage and decompression treatment,B group was given autologous PRP wound injection and follow-up on the basis of conventional treatment.After injection of PRP,platelet count,platelet recovery and application of PRP granulation tissue growth and wound healing were observed.Results The wound healing effect at the 7th,14th,21 st day between the two groups,and compared with the 1 st day,the differences were statistically significant (A group:10.28％,22.16％,43.25％,65.78％;B group:18.75％,37.58％,61.84％,80.26％;X2:7.895,8.934,10.231,9.076,all P ＜ 0.05).Two groups of patients had obvious difference in healing time,in B group the average healing time was (12.6 ± 5.2) d,which in was (17.3 ± 7.4) d,the difference was statistically significant (t =1.932,P ＜ 0.05).Conclusion The PRP can effectively promote the wound repair of soft tissue,to promote healing of chronic difficult wound healing,the curative effect is distinct,it is worthy of clinical popularization and application.

Objective: To evaluate the efficacy and safety of overlapping biodegradable polymer sirolimus-eluting stents (EXCEL) in treating the patients with diffuse long coronary lesions (total stent length for per lesion>60 mm).
Methods: A total of 71 patients with diffuse long coronary lesions with overlapped EXcellstents implantation in our hospital from 2010-08 to 2012-05 were retrospectively studied. The average age of patients was (62.85 ± 10.26) years and 74.56%with male gender. The clinical endpoints were the major adverse cardiac events (MACE) at in-hospital time and at 2-year follow-up period.
Results: The average target lesion was implanted (2.61 ± 0.52) stents, the mean stent diameter was (3.21 ± 0.35) mm and the length was (73.34 ± 13.11) mm. The in-hospital MACE rate was 4.23%, the 2-year target vessel revascularization and MACE rates were 9.86%and 18.31%respectively. Cox regression analysis indicated that smoking (HR 12.102, 95%CI 1.460-100.309, P=0.021), previous history of MI (HR 11.948, 95%CI 1.144-124.726, P=0.038) and previous history of PCI (HR 0.097, 95%CI 0.010-0.990, P=0.049) were the independent risk factors of out of hospital MACE occurrence.
Conclusion: EXcellstent implantation was safe and effective for treating the patients with diffuse long coronary lesions, the long term follow-up study revealed that there was the increased risk for MACE and target vessel revascularization.

Objective To investigate the clinical characteristics of the human Adenovirus (HAdv) infections in allogeneic hematopoietic stem cell transplantation ( allo-HSCT) patients and explore the clinical significance of HAdv monitoring .Methods A total of 845 cases underwent allo-HSCT were included retrospectively in Perking University People′s Hospital from October 2012 to August 2014.Peripheral blood HAdv load were monitored twice weekly within 100 days after allo-HSCT, or whenever necessary quantitatively by real-time PCR. Meanwhile, other clinical samples such as stool , urine, and bronchoalveolar lavage fluid ( BLAF ) were also detected qualitatively whenever necessary .The follow-up period was at least six months after allo-HSCT.All clinical data were collected and analyzed .Results The total positive rate of HAdv was 3.4% ( 29/845 ) .The incidence of HAdv infection was higher in children [3.8%(6/155), <18y] than that of adults [3.3%(23/690),≥18y].HAdv infection diagnosed within 100 days after allo-HSCT accounted for 72.4%(21/29) of the total number of positive cases .There were 19 cases detected positive in peripheral blood , 16 cases in stool , 9 cases in urine , and 1 cases in BLAF , respectively.One patient was positive in peripheral blood , stool and urine.The overall median time of HAdv was 69 (13-189) d.The median time was 56 (53 -144) d in stool ,which was earlier than that of in peripheral blood , urine and stool.Among 29 cases of HAdv positive patients , 17 patients were coinfected with Cytomegalovirus(CMV) and 11 casess with Epstein-Barr virus(EBV).Twenty-five cases of HAdv were diagnosed with acute graft-versus-host disease(aGVHD) before HAdv infection, and 4 cases were diagnosed with chronic graft-versus-host disease ( cGVHD ) . The most common clinical manifestation was HAdv enteritis (14 cases), followed by hemorrhagic cystitis (7 cases).Two cases complicated with multiple organ injury ( >2 ) clinically, 1 cases with pneumonia.There were 8 cases of death at the end of follow-up.Conclusions HAdv is an important pathogen causing infection in patients after allo-HSCT. The infenction is characterized with multiple organ involvement .CMV and EBV coinfection is common .HAdv monitoring was of great significance in allo-HSCT patients.