Objective To improve the diagnosis and treatment of infective endocarditis(IE)by exploring its causes,pathogenic microorganism and clinicsI characteristics.Methods The clinical data of 120 IE patients treated in Peking Union Medical College Hospital from October 1997 to September 2007 were analyzed retrospectively.Results Of the 120 consecutive cascs diagnosed as IE according to the Duke's new criteria,79 were male and 41 female with a average age of(43.2±16.7)years old.Twelve cases were prosthetic valve endocarditis(PVE)and 108 cases native valve endocarditis(NVE)and there were no previously known heart diseases in 29 of the cases.Seventy-nine of the 108(73.1%)NVE patients had basic cardiac abnormalifies before IE diagnosis,such as congenital cardiovascular disease(30 cases),idiopathic mitral valve prolapse(23 cases)and rheumatic heart disease(11 cases).Fever(100.0%),anemia(54.2%)and embolism(48.3%)were the most common clinical manifestations in the IE development.Of the 83 patients who had a positive blood culture result,Streptococcus vividaus(51.8%)was the most common isolated microorganism.Conclusions Congenital cardiovascular diseases and idiopathic mitral valve prolapse are the two most commonly heart diseases in IE.Blood culture and echocardiogram should always be done to exclude IE,especially presenting with fever of unknown reasons.

Objective To explore the value of spectral imaging technique in dual-energy CT in differential diagnosis of the metastatic lymph nodes in thyroid carcinoma,squamous cell carcinoma metastatic lymph nodes and lymphoma in the neck.Methods In 30 patients with pathologically confirmed with a total of 79 cervical lymph nodes enlargement which were using dual energy scan .Then observed the change trend of the spectrum curve and comparison the three kinds of lymph node energy spectrum curve’s slope.Results In the 79 lymph nodes,the metastatic lymph nodes in thyroid carcinoma were twenty-three,squamous cell carcinoma metastatic lymph nodes were twenty-four and lymphoma were thirty-two.From 60 to 180 keV,with the increase of keV values,the three kinds of malignant lymph nodes of the corresponding CT value decreasing and the higher the keV value,the CT value decrease magnitude was small,and the spectrum curve was〞drop type〞.The slope spectrum curve of the metastatic lymph nodes of thyroid carcinoma in arterial phase and parenchymal phase were maximum,which were 1.23±0.41 and 0.85±0.33,respectively.The slope spectrum curve of lymphoma in arterial phase and parenchymal phase were least,whcih were 0.40±0.16 and 0.47 ±0.09.The slope spectrum curve of the squamous cell carcinoma metastatic lymph nodes in arterial phase and parenchymal phase were 0.88±0.10 and 0.62±0.28.The energy spectrum curve slope of the three kinds of malignant lymph nodes have statistical significance.Conclusion The energy spectrum curve slope of arterial phase and parenchymal phase has some significance lymph node metastasis of in thyroid carcinoma,the metastatic lymph nodes and lymphoma in the neck.

Objective To set up a technical platform of reverse genetics based on the 8 plasmid.virus rescue system of cold-adapted influenza virus strain. Methods The cold-adapted, temperature sensitive, live attenuated influenza virus strain A/AnnArbor/6/60(H2N2)was chosen as the master donor virus(MDV)for rescue research,and its six internal gene fragments PB2,PB1,PA,NP,M and NS were artificially synthesized. Meanwhile, five amino acid mutations have been introduced as tags. Six fragments were ligated with modified pAD3000 for the construction of rescue plasmid. Six transcription/expression plasmids(pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,and pMDV-A-NS)were obtained, and their sequences were accurate. Results The reassorted virus named as rMDV-A contains HA and NA gene segments derived from PR8 strain along with six gene segments,PB2,PB1,PA,NP,M and NS,from MDV. The COS-1 cells were co-transfected with eight recombinant plasmids. The results showed that a cold-adapted, attenuated reassortant influenza A virus with hemagglutination activity was rescued successfullv bv"6+2" combination of MDV and PR8, and the allanotoic fluid of the injected eggs gave a posigenes of A/AA/6/60 used as backbone has provided experimental materials for further research on the gene function and novel vaccine candidate of cold-adapted, attenuated human influenza virus.

Six gene segments,PB1,PB2,PA,NP,M and NS,were fully synthesized which derived from the master donor virus (MDV),cold-adapted(ca),temperature sensitive(ts),live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A).Meanwhile,five amino acid substitutions (PB1-391E,58lG,661T,PB2-265S,NP-34G) were artificially altered by human intervention.HA and NA fragments derived from the 2006-2007 circulating strain A/New Caledonia/20/99 (H1N1).Eight fragments were ligated with modified pAD3000 for rescue plasmid construction.Eiight transcription/expression plasmids were named as pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,pMDV-A-NS,pMDV-A-HA,pMDV-A-NA,respectively.The COS-l cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the CDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1),the results showed that cold-adapted,attenuated reassortant influenza A virus Was rescued successfully.Titers of a reassorted influenza A virus in embryonated chicken eggs mnged from 1:29to l:210.The rescue system of six intemal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted,live attenuated human influenza virus.

Six gene segments, PB1, PB2,PA, NP, M and NS, were fully synthesized which derived from the master donor virus(MDV), cold-adapted(ca),temperature sensitive(ts), live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A). Meanwhile, five amino acid substitutions (PB1-391E, 581G, 661T, PB2-265S, NP-34G) were artificially altered by human intervention. HA and NA fragments derived from the 2006～2007 circulating strain A/New Caledonia/20/99 (H1N1). Eight fragments were ligated with modified pAD3000 for rescue plasmid construction. Eight transcription/expression plasmids were named as pMDV-A-PB2, pMDV-A-PB1, pMDV-A-PA, pMDV-A-NP, pMDV-A-M, pMDV-A-NS, pMDV-A-HA, pMDV-A-NA, respectively. The COS-1 cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the cDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1), the results showed that cold-adapted, attenuated reassortant influenza A virus was rescued successfully. Titers of a reassorted influenza A virus in embryonated chicken eggs ranged from 1∶29 to 1∶210. The rescue system of six internal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted, live attenuated human influenza virus.

A cold-adapted (ca), temperature sensitive (ts), live attenuated influenza B virus strain B/Ann Arbor/1/66 was chosen for influenza virus rescue research, in which six internal gene segments, PB1, PB2, PA, NP, M, NS, were fully synthesized and nine amino acid substitutions were artificially alter by human intervention. The resultant B/Ann Arbor/1/66 plasmids were named as pAB121-PB1, pAB122-PB2, pAB123-PA, pAB124-HA, pAB125-NP, pAB126-NA, pAB127-M and pAB128-NS, respectively. A recombinant influenza A virus was previously generated entirely from cloned cDNA. An infectious recombinant influenza B virus was generated here, and designated as rMDV-B, by plasmid-based reverse genetics. The rMDV-B virus contained HA and NA genes from an epidemic influenza B vires strain B/Malaysia/2506/2004 in the background of internal genes derived from influenza B virus strain B/Ann Arbor/1/66. HA titer of rMDV-B in MDCK cells and embryonated chicken eggs ranged from 1 : 64 to 1 : 512. The results may allow an effective live influenza B vaccine to be produced from a single master strain, providing a model for the design of future live human influenza vaccines.

Objective To explore the clinical diagnosis and treatment of hyperthyroidism patients coexisted thyroid carcinoma.Methods A retrospective analysis was made in 15 cases with hyperthyroidism patients coexisted thyroid carcinoma who were operated from Nov.2008 to Dec.2017 in Department of General Surgery of Jian'an District Peoples' Hospital.Results The incidence of hyperthyroidism patients coexisted thyroid carcinoma was 8.19％(15/183) in our study.All 15 patients included 12 papillary thyroid carcinoma,1 follicular thyroid carcinoma,1 medullary thyroid carcinoma,and 1 follicular papillary carcinoma.Among them,papillary thyroid microcarcinoma accounted for 66.67％(10/15).All 15 postoperative patients were followed up and the mean time was 13.25 months.Neither recurrence nor mortality occurred during the period.Conclusions Hyperthyroidism patients coexisted thyroid carcinoma have no characteristic clinical manifestations or specific diagnosis indicators.The prognosis can be good through strengthening awareness,improving vigilance,comprehensive analysis and surgical treatment.

Objective: To analyze the clinical characteristics and prognosis of 34 cases of acute myeloid leukemia (AML) with FLT3 internal tandem duplication (FLT3-ITD) and MLL gene rearrangement. Methods: The clinical data of 34 AML patients with FLT3-ITD and MLL gene rearrangement was compared and analyzed for the therapeutic efficacy, prognostic factors when treated with chemotherapy, chemotherapy combined with targeted therapy or allogenic hematopoietic stem cell transplantation (allo-HSCT). Results: Of the thirty-four cases with median age 41 (4-71) years old, 63.6% presented with white blood cells (WBC) greater than 30×10⁹/L, 39.4% greater than 50 × 10⁹/L respectively on admission. M₅ (35.3%) made up the highest proportion. The cytogenetic abnormality reached 61.8%, of which the complex cytogenetic abnormality accounted for 11.8%. Eleven patients (32.35%) had both FLT3-ITD and MLL gene abnormalities. In addition to FLT3 and MLL abnormalities, 23 patients (67.6%) had one or more other gene abnormalities (multiple gene abnormalities). Of the 34 cases, 29.4% patients went into complete remission (CR) after two courses of chemotherapy. 20.6% (7 patients) went into CR after 3 or more courses of chemotherapy. The rate of early relapse in the CR group was 52.9%. Patients with WBC>50×10⁹/L or multiple gene abnormalities had a lower remission rate (7.7%, 5.4%) after two courses of chemotherapy. CR rate for the patients with more than three gene abnormalities was 0. The total 2-year overall survival (OS) in the 34 patients was 28.8% (95% CI 13.5%-46.0%) and the disease-free survival (DFS) was 27.1% (95% CI 12.5%-44.0%). Of the 18 patients treated with chemotherapy alone or chemotherapy combined with targeted therapy, 17 cases died within 2 years and 1 lost follow-up after giving up treatment. For the 16 patients received allo-HSCT, the 3-year OS was 43.4% (95% CI 13.7%-70.4%) and DFS 42.7% (95% CI 13.4%-69.7%). Conclusion AML patients with FLT3-ITD and MLL gene rearrangement often presented with M₅, accompanied by hyperleukocytosis, cytogenetic or multiple gene abnormalities. Those patients were observed to have low response rate and high early relapse when treated with chemotherapy without allo-HSCT. Patients had multiple gene abnormalities may be an important poor prognostic factor. Allo-HSCT is an effective treatment which could significantly improve the prognosis and survival of AML patients with FLT3-ITD and MLL gene abnormalities.

Objective To prepare and identify CS-PLGA microspheres carrying bilayer antigen protein.Methods Porous microspheres were prepared by emulsion polymerization combined with solvent evaporation.The antigen was loaded in a water bath at 4 ℃,and the antigen was promoted to enter the microspheres by physical diffusion media-ted by antigen concentration gradient and combined with the outer surface of the microspheres by electrostatic ac-tion,forming an inner and outer double-layer antigen carrier.Then,according to the glass transition temperature of PLGA material,the pores on the surface of porous microspheres were promoted to heal at 48 ℃ to form a closed microsphere preparation,and then the obtained microsphere preparation was suspended with chitosan solution for cationic modification to reverse the negative potential on the surface of microspheres.In this study,the morphology,particle size distribution and potential changes of microspheres were detected by scanning electron microscope and dynamic light scattering particle size analyzer.In addition,sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)was used to identify whether the antigen was loaded into the microsphere preparation.The fluorescent labeled BSA and fluorescent labeled PLGA materials were used to observe the distribution of the antigen after load-ing by laser confocal microscope.The encapsulation efficiency and drug loading rate of the microsphere preparation were detected by BCA method.Results The results of scanning electron microscope and optical microscope showed that the porous microspheres had good pore formation,the particle size was(73.94±0.81)nm,and the Polydisper-sity index was 0.038±0.004.Zeta potential changed from negative to positive,which indicated that chitosan had been successfully coated on the surface of microspheres.SDS-PAGE,laser confocal microscope and other detection methods confirmed that BSA had been successfully carried.The encapsulation rate of porous microspheres was(3.01±0.04)％and the drug loading rate was(1.50±0.02)％after detection by micro BCA kit.Conclusion CS-PLGA preparation carrying bilayer antigen was successfully prepared,which provided a new idea for the subse-quent study of sustained and controlled release preparations.

Objective:To evaluate the efficacy and safety of anti-human T lymphocyte porcine immunoglobulin (P-ATG) with recombinant human tumor necrosis factor-α receptor Ⅱ:IgG Fc fusion protein (rhTNFR∶Fc, Etanercept) on grade Ⅲ/Ⅳ acute graft-versus-host disease (aGVHD) after allogenic hematopoietic stem cell transplantation (allo-HSCT) .Methods:Thirty-five patients with Grade Ⅲ/Ⅳ aGVHD who received P-ATG with etanercept therapy after allo-HSCT were retrospectively analyzed. P-ATGs (5 mg·kg -1·d -1) were administrated for 3 to 5 days, and then 5mg/kg was sequentially administrated, QOD to BIW. Etanercepts were administrated 25 mg, twice a week (12.5 mg, BIW for pediatric patients) . Results:Among the 35 patients with grade Ⅲ/Ⅳ aGVHD, 21 were males and 14 females, with a median age of 10 (3-54) years. A total of 19 cases of acute myeloid leukemia, 13 of acute lymphoblastic leukemia, 1 of severe aplastic anemia, 1 of myelodysplastic syndrome, and 1 of mixed phenotypic acute leukemia were noted. The overall response (OR) rate of P-ATG with etanercept was 85.7% (30/35) , with complete response (CR) and partial response (PR) rates of 34.3% (12/35) and 51.4% (18/35) , respectively, on day 28. The OR rate of grade Ⅲ aGVHD group was higher than of grade IV aGVHD group [100% (19/19) vs. 68.8% (11/16) , P=0.004]. On day 56, the OR rate became 77.2% (27/35) , with CR and PR rates of 62.9% (22/35) and 14.3% (5/35) , respectively. The OR rate of grade Ⅲ aGVHD group was also higher than of grade Ⅳ aGVHD group [89.5% (17/19) vs. 62.5% (10/16) , P=0.009]. Thirty-five patients had no adverse effects such as fever, chills, and rash during the P-ATG infusion, and no obvious liver and kidney function damage was observed after treatment. The main treatment-related complication was infection. The reactivation rates of CMV and EBV were 77.1% (27/35) and 22.9% (8/35) , respectively, and the bacterial infection rate was 48.6% (17/35) . With a median follow-up time of 13 (1-55) months after HSCT, the 1-year and 2-year OS rates were (68.1±8.0) % and (64.3±8.4) % , respectively. The 1-year OS rate of grade Ⅲ aGVHD group was superior to grade Ⅳ aGVHD group [ (84.2±8.4) % vs. (47.6±13.1) % , χ2=3.38, P=0.05]. Conclusion:This study demonstrated that P-ATG with etanercept was effective and safe in treating grade Ⅲ-Ⅳ aGVHD after allo-HSCT.