Objective To investigate the research quality of present medical graduates and propose suggestions for improvement.Method A self-designed questionnaire entitled Investigating Questionnaire for Research Quality Analysis of Medical Graduates in Peking Union Medical College Hospital was distributed to the clinical and academic medical graduates in the hospital from July to August 2015.Among 276 collected questionnaires,270 were validated as effective.SPSS 18.0 software was used to statistically describe the result and to perform t test on different subgroups.Results The research quality average of the surveyed hospital was 10.28.Results revealed that in regard of the three aspects of research quality,research consciousness ranked first,followed by the research ability,while scientific spirit was the weakest.Academic medical graduates showed significantly higher scores than clinical graduates in terms of total research quality and every single aspect (P values less than 0.05).Conclusions To better cultivate the academic leaders in medical research,it is necessary to strengthen the research training for the graduates,practice their scientific thinking especially the clinical graduates,reinforce tutors' guidance,and promote communication and collaboration.

OBJECTIVE To investigate the incidence and pathogens of infection in 1659 consecutive cases in single center hematological unit.METHODS The incidence,pathogen,and outcome of infection in 1659 hospitalized patients with hematological disorders from 1999 to 2006 were retrospectively analyzed.RESULTS The overall incidence of infection was 24.4% according to the person-times of hospitalization,which included 22.1% of nosocomial infection and 2.3% of community acquired infection.Most of the pathogenic bacteria of the nosocomial infection were Gram-negative.The most common bacteria in the sputum samples included Enterobacter cloacae(19.3%) and Pseudomonas aeruginosa(14.8%).The most common bacteria in the blood samples were coagulase negative Staphylococcus(CNS,39.3%),the next was Pseudomonas aeruginosa and Escherichia coli.There were 4.21% bacteria resistant to most of antibiotics in nosocomial infection.There were 114 fungi isolated.Candida albicans was accounted for 35.1%.The mortality due to nosocomial infection was 7.4%.CONCLUSIONS The patients in hematology ward are susceptible to infection.The pathogens of nosocomial infection are most likely G-bacteria.Some bacteria are resistant to almost all antibiotics.The incidence of fungal infection is increasing in the near 8 years.

Objective To explore the cytogenetic characteristics of acute myeloid leukemia(AML) patients.Methods The karyotype analysis was performed in 178 AML using the short-term culture of bone marrow cell and G-banding technique.Results Among the 178 patients,171 had enough metaphases for analysis and 128(74.9%)had clonal karyotypic abnormalities.Twenty-seven patients were secondary to myelodysplastic syndrome (MDS-AML),with 25 (92.6%) patients carrying clonal karyotypic abnormalities.Among the remaining 144 patients of de novo AML,103(71.5%)had clonal karyotypic abnormalities.The rate of abnormal clonal karyotype was higher in MDS-AML than that of de novo AML (P=0.021).Among the 171 patients,41(24.0%)were in favorable risk group,80(46.8%)in intermediate risk group and 50(29.2%)in adverse risk group.t(15;17)was the most common chromosomal aberration.The maiority intermediate risk chromosomal aberration was;normal karyotype.The most common cytogenetic abnormality among adverse group was a complex karyotype.Adverse cytogenetic aberrations,such as -5/5q-,-7/7q-,frequently occurred in conjunction with one another as part of a complex karyotype.Totally 75 patients were 60 years or older,among them,16.0%were in favorable risk group,48.0%in intermediate risk group and 36.0%in adverse risk group.Among 96 younger patients,30.2%were in favorable risk group.45.8%in intermediate risk group and 24.0%in adverse risk group.The rate of favorable risk chromosomal aberration was lower in elder patients than in younger(P=0.03 1).The rate of adverse risk chromosomal aberration and the rate of monosomal karyotype were higher in MDSAML than in de novo AML patients(P<0.001).Conclusions The most common favorable,intermediate and adverse chromosomal aberrations were t(15;17),normal karyotype and complex karyotype respectively.The karyotype was poor in MDS-AML and elder AML patients.

Objective To explore the clinical characteristics,therapy reactions and prognosis of the elderly patients with idiopathic thrombocytopenic purpura(ITP). Methods A total of 43elderly ITP patients(age≥60 years old)including 16 men and 27 women were reviewed and further followed up for 1 month to 15 years. Results Until June 2007,35 elderly ITP patients survived,platelet counts were sustained(30-50)×109/L in 7 cases,but no significant bleeding was found.Thirty-six patients had adrenocorticosteroid therapy first, 25 patients were sensitive to adrenocorticosteroid therapy,4 patients underwent splenectomy,and 3 patients achieved a normal platelet count. Immunosuppressive agents(vinscristine,cyclophosphamide, azathioprine and Cyclosporin A)treatments were held in 5 6 case-times,Cyclosporin A and azathioprine were more effective than vinscristine and cyclophosphamide.The refractory rate was 13.9%.One patient progressed to monoclonal gammopathy of unknown significance and 1 to lymphoma.Eight patients died.1 patient died of brain bleeding after trauma,3 patients died of malignant neoplasm,4 patients died of heart failure induced by infection. Conclusions The clinical features of elderly ITP patients are atypical.the mortal bleeding in them was rare,treatment strategy should be individualized tO each elderly patient.

Objective To explore the karyotype distribution in elderly patients with acute leukemia (AL) and compare the prognostic characteristics of karyotype by age grouping.Methods Chromosomal karyotypes were analyzed in 215 cases with AL using the short-term culture of bone marrow cells and G-banding technique.Results There were 202 cases with enough mitosis for analysis and 149 cases(73.8％)with abnormal clone in 215 patients with AL.The rates of abnormal clone were 73.0％ (27/37),74.4％(64/86) and 73.4％ (58/79) in patients aged ≤30,31-59 and ≥60 years,respectively,and no difference were found among age groups (P=0.982).Among 171 patients with acute myeloid leukemia (AML) with detected mitosis,there were 41 better-risk cases (24.0 ％) with most frequent aberration of t(15;17) accounting for 65.9 ％,80 intermediate-risk cases (46.8 ％ ) with principal of normal karyotype accounting for 53.8 ％,and 50 poor-risk cases (29.2 ％)with complex karyotype occupied by 84.0％.The karyotype percentage of better-risk,intermediaterisk and poor-risk were 50.0％,36.4％ and 13.6％ in patients aged ≤30 years,24.3％,48.7％ and 27.0％ in aged 31-59 years,and 16.0％,48.0％ and 36.0％ in aged ≥ 60 years,respectively.The rate of better-risk karyotype was higher in patients aged ≤30 years than the other two groups (P=0.021and P=0.001) and the ratio of poor-risk karyotype higher in patients aged ≥ 60 years than in patients aged ≤30 years (P=0.046).Among 29 patients with acute lymphoblastic leukemia (ALL),10 cases had poor-risk and 19 cases had intermediate-risk karyotype.Conclusions Karyotype analysis provides an important basis for risk assessment and the rate of poor-risk karyotype may increase with the ageing in patients with AML.

Objective To explore the outcome of monoclonal gammopathy of undetermined significance (MGUS). Methods The data from 14 MGUS patients in our hospital including clinical features, outcome and change of M protein concentration were analyzed retrospectively. Results The MGUS didn't have the clinical manifestations of multiple myeloma (MM), the time of outcome from MGUS to MM was about 4-20 years (mean time, 10 years). The most types of MM were IgA and IgG, 6 cases were IgA type, 6 cases were IgG type and 2 cases were light chain type. The concentration of immune globulin in general showed an upward trend year by year. A few showed fold lines ascend. Conclusions The elevated monoclonal immunoglobulin may develop into MM after many years. We must follow up frequently to avoid error diagnosis and missed diagnosis.

In this new era of the genome, the complete sequences of various organisms (from the simplest to the most complex such as human) are now available, which provides new opportunities to study biology and to develop therapeutic strategies. But the paucity of research tools that manipulate specific genes in vivo represents a major limitation of functional genomic studies. In nature, the expression of genes is regulated at the transcriptional level primarily by proteins that bind to nucleic acids. Many of these proteins, which are termed transcription factors, are typically consist of two essential yet separable modules: DNA-binding domain (DBD) and effector domain (ED). Attempts to control the gene expression by artificial transcription factors are based on the application of this rule. Among the many naturally occurring DNA-binding domains, the Cys2-His2 zinc-finger domain has demonstrated the greatest potential for the design of novel sequence-specific DNA-binding proteins. Each zinc finger domain, which comprises about 30 amino acids that adopt a compact structure by chelating a zinc ion, typically functions by binding 3 base pairs of DNA sequence. Several zinc fingers linked together would bind proportionally longer DNA sequences. Ideally, these artificial DNA binding proteins could be designed to specifically target and regulate one single gene within a genome as complex as that found in human. Such proteins would be powerful tools in basic and applied research.
DNA ; chemistry ; genetics ; metabolism ; DNA-Binding Proteins ; metabolism ; Gene Expression Regulation ; Humans ; Transcription Factors ; chemistry ; genetics ; metabolism ; Zinc Fingers ; genetics ; physiology

Transcriptions are regulated by transcription factors. Natural transcription factors usually consist of at least two functional domains: a DNA-binding domain and an effector domain. According to this, novel artificial transcription factors are designed to up or down regulate transcription and expression of a target gene. The Cys2-His2 zinc finger domain is a DNA-binding module that has been widely used as the DNA-binding domain in artificial transcription factors. Each zinc finger domain, which comprises about 30 amino acids that adopt a compact structure by chelating a zinc ion, typically functions by binding 3 base pairs of DNA sequence. Several zinc fingers linked together would bind proportionally longer DNA sequences. According to the "bipartite complementary" library strategy, a pair of zinc finger phage display libraries were constructed. After construction of the libraries, a 9bp sequence (5'-GCAGAGGCC-3') on the promoter of SV40 was chosen as a target for next step. After parallel selection, PCR amplification, desired fragments recovery, re-ligation, and additional rounds of selection, phage enzyme-linked ELISA experiments were performed to identify specific binding clones displaying the zinc fingers with predetermined sequence-specificity to our target sequence. Then two clones with strong ELISA signals were chosen to be tested for binding both to its full target site (5'-GCAGAGGCC-3') and to sites containing single transition mutations. The binding specificity of one of the two clones (clone 3) was shown to be fairly good. The three-finger DNA-binding domain targeted to SV40 promoter, that is, zinc finger sequences on clone 3, was fused to KOX1 suppression domain KRAB and cloned into pcDNA3.1 (+) (which expression product was artificial transcription factor). The zinc fingers (which expression product was the DNA-binding domain of artificial transcription factor) and KRAB domain only (which expression product was effector domain of artificial transcription factor) were also cloned separately into the same expression vector. All constructs contained an N-terminal nuclear localization signal. Every of the vectors (including pcDNA3.1 (+) without inserting sequences) were cotransfected with pGL3-Control and pRL-TK and the activity of luciferase was used to indicate the function of product from transfected expression vectors. Our artificial transcription factor was proved to repress the expression of reporter gene efficiently,while with only DNA-binding domain or effector domain the repression was not remarkable. By adding different effector domains and changing the DNA-binding domain, artificial transcription factor would have a wide range of potential applications.
Enzyme-Linked Immunosorbent Assay ; Genes, Synthetic ; genetics ; physiology ; Models, Theoretical ; Peptide Library ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Transcription Factors ; chemical synthesis ; chemistry ; metabolism ; Zinc Fingers ; genetics ; physiology

AIM: To evaluate the anatomic and visual outcomes of 25-gauge vitrectomies combined with air tamponade for the treatment of idiopathic macular hole (IMH).METHODS: Thirty eyes of 27 patients with IMH were included in this prospective interventional study.All patients underwent 25-gauge pars plana vitrectomy (PPV) combined with phacoemulsification and air tamponade.Best corrected visual acuity (BCVA, logMAR), perimetry and multifocal electroretinography (mfERG) were conducted before and after the operation.Anatomical changes were evaluated with optical coherence tomography (OCT).RESULTS: The macular holes closed successfully in 28 eyes after the primary vitrectomy.The mean BCVA improved from 0.72±0.22 logMAR preoperatively to 0.29±0.18 logMAR postoperatively (P<0.001).In the visual field of central 10°, the average mean deviation (MD) decreased from-3.59±1.83 dB preoperatively to-2.51±1.36 dB postoperatively (P<0.001) and the average pattern standard deviation (PSD) decreased from 1.86±0.68 dB preoperatively to 1.33±0.32 dB postoperatively (P=0.001).The retinal response densities of mfERG in the foveal and perifoveal area increased significantly, and implicit times of rings 4-6 prolonged significantly (P<0.05).The symptom duration and baseline N1 amplitude densities at ring 1 had a significant impact on postoperative BCVA (P<0.001, P=0.001, respectively).CONCLUSION: The 25-gauge PPV and air tamponade with 1 day prone positioning produce favorable anatomic and functional outcomes.

BACKGROUND: Osteoporosis is a genetic disease associated with many enes. To date, the genes that regulate bone mass are incompletely defined.OBJECTIVE: To investigate the relationship between polymorphisms of steoprotegerin (OPG) gene promoter with bone mineral density (BMD) in remenopausal and postmenopausal women.DESIGN: Prospective study.SETTING: Peking Union Medical College Hospital.PARTICIPANTS: In July 2002, 495 Han nationality women selected from Peking Union Medical College Hospital were non-related volunteers and gave their informed consent prior to the study, which included 306 premenopausal women aged 20-39 years, 189 postmenopausal women aged 50-84 years.METHODS: ① BMD measurement: BMD was measured at the Lumbar Spine and Femoral Neck, trochanter, Ward's triangle by dual-energy X-ray absorptiometry. ② Genotyping: Whole blood genome DNA was extracted by QIAGEN DNA extraction kit. The PCR product and the result of endonuclease digest were confirmed by sequencing (Bioasia Biotechnology,Shanghai, China). The impact of the polymorphisms on BMD was also investigated using multiple Logistic regression.MAIN OUTCOME MEASURES: ① Distribution of OPG genotypes and the relationship with BMD. ② Association between OPG polymorphisms and osteoporosis.RESULTS: All 495 subjects were involved in the final analysis. ① These polymorphisms were in Hardy-Weinberg equilibrium (χ2= 0.056 -0.222, P＞ 0.05). The frequencies of genotypes of these subjects were as follows: AA (70.1%), AG (26.9 %), GG (3.0 %) for 163A→G polymorphism; TT (71.3 %), TG (25.9 %), GG (2.8 %) for 245T→G polymorphism. BMD was lower in premenopausal women with GG +AG genotype than AA genotype for 163A→G polymorphism, so did GG+TG genotype than TT genotype for 245T→G polymorphism. But there was no significant difference. BMD was lower in postmenopausal women with AG+GG genotype than AA genotype for 163A→G polymorphism at Lumbar Spine 2-4, Femoral Neck, Ward's triangle and Trochanter (P ＜ 0.05). For 245T→G polymorphism, BMD of postmenopausal women with TG+GG genotype was lower at Femoral Neck,Ward's triangle and Trochanter than TT genotype (P ＜ 0.05). For 245T→G polymorphism, BMD of postmenopausal women with TG+GG genotype was lower at Femoral Neck, Ward's triangle, and Trochanter than TT genotype (P ＜ 0.05). ② Age, weight, height, years since menopause, and 163A→G/245T→G genotypes were sewed as covariates. AG+GG genotype was contributed to low BMD at Lumbar Spine 2-4 and Ward's triangle (OR =2.045, OR=2.956, P ＜ 0.05, 95% CI 1.05-6.7). TG+GG genotype was risk factor for osteoporosis at Lumbar Spine 2-4, Ward's triangle,and Trochanter (OR=2.059, OR=2.859, OR=2.123, P ＜ 0.05, 95% CI 1.04-6.5).CONCLUSION: BMD was lower in postmenopausal women with the variant G allele for 163A→G and 245T→G polymorphisms at Femoral Neck,Ward's triangle, and Trochanter. The variant allele G may associate with lower BMD in postmenopausal women.