Objective To evaluate the values of the combined eGFR and qualitative determination of urine albumine on in both during and 3 years after hospitalization prognosis of after PCI.Methods One thousand and five PCI patients were divided into 4 groups by level of eGFR and qualitative determination of urine albumine.A group:eGFR ＜ 60 ml/min,urine albumine negative ( n =34 ) ; B group:eGFR ＜ 60 ml/min,urine albumine positive ( n =34) ; C group:eGFR ≥60 ml/min,urine albumine negative ( n =797 ) ; D group:eGFR ≥60 ml/min,urine albumine positive (n =140).The levels of serum creatinine and urine albumine of PCI patient were measured,and then compare body mass index,smoking,hypertension,hyperlipaemia,diabetes mellitus,previous MI,perioporative PCI and etc.All patients were followed up from 30 d to 3 years.The prognostic facters of PCI patients were analyzed by COX proportional hazards models.Kaplan-Meier survival analysis was used to compare survival curves between the four groups stratified by level of eGFR.The log rank statistic was used to test for differences between groups.Results By multivariate COX regression adjustment for body mass index,smoking,hypertension,hyperlipaemia,diabetes mellitus,previous MI,perioperative PCI,the relative risk (RR) of albuminuria for cardiac events was 2.006 ( 95％ CI:1.020 - 3.947,P ＜ 0.05 ),perioperative PCI 3.375 ( 95％ CI:2.106 - 5.410,P ＜0.05 ).The cardiac events rate of B group with PCI after 1,2 and 3 year was 20.59％,2.94％,2.94％respectively,have statistical significant(x2 =8.774,P ＜ 0.05 ).The cardiac events rate of the first year in A,B,C,D groups was 5.89％,20.59％,4.01％,2.86％ respectively.The different cardiac events rate among four groups had statistical significant(x2 =22.050,P ＜0.01 ).B group cardiac events rate of the first year was significant higher than C,D group,which had statistical significant (x2 =20.020,14.520,P ＜0.01 ).The survival rate of 3 year follow-up after PCI was 91.2％,73.5％,91.6％,93.6％ respectively in four group,the different survival rate among four groups have statistical significant(x2 =16.750,P ＜0.01 ).Conclusion For the identification and treatment of high incidence of cardiac events in patients undergoing PCI,eGFR and qualitative determination of urine albumine are simple and effect tools.

[Objective]To evaluate the clinical result after treatment on fracture of distal radius with different fixations of the wood splint/plaster splint/"T" model plate.[Method]There were 236 cases with the fracture of distal radius,there were divided three types of A,B,C according to the AO classification,70 cases in 236 cases were treated with close diaplasis and external wood splint fixation,55 in 236 cases with external plaster splint fixation,III in 236 cases with operation and internal "T" model plate fixation.Results were evaluated by replacement and function after treatment,and percentage of excellence was compared with three different fixation.[Result]236 cases were all followed-up,mean follow-up period was 14.6 months(6~20months).The excellence rate of diaplasis was not significantly different in A and B type by comparing three different fixation,the excellence rate of function with wood splint fixation and "T" type plate internal fixation was better than that of plaster splint fixation in A and B type,there were not significantly difference about the excellence rate of function with the external wood splint fixation and the internal "T" model plate fixation.The excellence rate of function and reduction with"T" type plate internal fixation were better than with plaster splint and wood splint in C type.[Conclusion]The author suggests open reduction with "T" type plate internal fixation about C type fracture,close reduction and external wood splint fixation about A and B type distal radius fracture.

Objective To evaluate the diagnostic value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) and conventional-transbronchial needle aspiration (C-TBNA) in mediastinal and lung hilar lesions. Method 301 cases of lung hilar and mediastinal lesions were selected from 2010 to 2016. Among them, 183 cases underwent TBNA, and the other 118 cases received EBUS-TBNA technology. During the research, the associations of diagnostic positive rate and complications were analyzed in order to explore the advantage and the value of EBUS-TBNA. Results The positive rates of EBUS-TBNA in central groups (2R, 4L, 4R, 7) were higher than in the peripheral groups (10R, 10L, 11R, 11L) (P < 0.05). When studying the lymph node group 2R, 4R and 7R, the positive rate of EBUS-TBNA is much more significant than conventional TBNA (P < 0.05); When biopsying at the lymph node group R4 and group 7, one needle positive rate of EBUS-TBNA were much more superior than TBNA (P < 0.05), the three needles cumulative positive rate of EBUS-TBNA almost reach the total positive rate(P > 0.05), an approving effect of puncture can be acquired; The accuracy and sensitivity of EBUS-TBNA in the diagnosis of lung hilar and mediastinal lesions were much better than conventional TBNA (P < 0.05), especially the diagnostic positive rate of EBUS-TBNA in benign diseases was higher (P < 0.05); The complications rates in both two technologies were not significantly different (P > 0.05), there were no severe complications during the operations in all cases. Conclusion EBUS-TBNA is useful in diagnosis of mediastinal and hilar lesions of unknown reason, and significant in diagnosis of bronchial and extrabronchial diseases. It is an efficiency and safe operation while further application studies are needed.

Objective To investigate the clinical results and related key points of surgical treatment for Hawkins Ⅲ talus neck fractures.Methods From March 2005 to March 2010,26 patients with Hawkins Ⅲ talus neck fracture were treated and 21 of them were followed,including 13 males and 8 females,with an average age of 37.6 years.The fractures occurred on the left side in 11 patients and on the right side in 10patients.The mechanism of injury included high falling injury in 13 patients,traffic accident injury in 7 patients and rolling down injury in 1 patient.Five cases were closed fractures and 6 cases were open fractures.A bilateral approach,the medial and lateral approaches,was used to perform the operation with cannulated screw fixation in emergency.After external fixation in functional position for 6 weeks without loading,ankle joints were allowed to take exercise with hinge brace and to bear partially basing on plain radiograph 12-16weeks later.The weight-bearing should be adjusted with follow-up.Functional results were assessed according to AOFAS (American Orthupaedic Foot and Ankle Society,AOFAS)score.Results The average duration of follow-up was 36.6 months (range,6-60 months).All fractures gained union and the average union time was 4.5 months.The average AOFAS score was 78.6.There were 4 cases in excellent results,10 in good,5 in fair and 2 in poor.The overall excellent and good rate was 67.8％.Traumatic arthritis occurred in 13 cases and avascular necrosis in 5 cases.Conclusion The effect of surgical treatment for Hawkins Ⅲtalus neck fracture via a bilateral approach is satisfactory.

Objective To evaluate the postoperative prognosis of the modification of diet in renal disease formula (MDRD) in coronary artery bypass graft surgery (CABG) in hospital or 4 years after hospitalization. Methods Two hundred and seventy-two CABG patients were divided into 3 groups according to the levels of estimated glomerular filtration rate (eGFR) including 35 cases in eGFR < 60 ml/min group, 119 cases in 60 ≤ eGFR < 90 mL/min group and 118 cases in eGFR ≥90 ml/min group. The prognostic factors of CABG patients were analyzed by COX proportional hazards models. Kaplan-Meier survival analysis was used to compare survival curves among the three groups stratified by eGFR levels. The Log-rank statistic was used for comparing between groups. Results By multivariate COX regression adjustment for body mass index, smoking, hypertension, hyperlipaemia, diabetes mellitus, previous MI, perioperative PCI and etc. , the relative risk (RR) of the increasing age for cardiac events was 1.077(95% CI 1.002-1.158,P =0.044). RR of left ventricular ejection fraction (LVEF) was 0.005(95% CI 0.000-0.456,P =0.022). RR of eGFR was 0.968(95% CI 0.948-0.988,P =0.002). The survival rate in the first, second, third and fourth year were same in every group. The survival rate of group with eGFR < 60 ml/min, 60 ≤ eGFR < 90 ml/min and eGFR≥ 90 mL/min was 76.4%, 93.1%, and 96.6%. The survival rates among three groups were statistically significant. In the survival curve of 4 year follow-up after CABG, the survival rate of group with eGFR < 60 mL/min was lower than that of 60 ≤ eGFR < 90 ml/min group and eGFR ≥ 90 ml/min group. Conclusions The preoperative eGFR is an independent risk factor in evaluating cardiac events in hospital and after hospitalization. It has a higher prognosis value in patients undergoing CABG.

Objective To explore the relationship between the HLA-G 14 bp insertion/deletion polymorphism and the infection of Epstein-Barr virus(EBV) for children.Methods The study genotyped HLA-G 14 bp insertion/deletion polymorphism of 102 infectious mononucleosis children and 165 normal controls by PCR-PAGE,detected the plasma sHLA-G level of 51 infectious mononucleosis children and 146 normal controls by ELISA.Results A significant difference was observed for the frequencies of the HLA-G 14 bp genotype between the two groups( x2 =6.742,P=0.034 ),and a significant difference was also observed for the 14 bp allele frequencies between the two groups( x2 =6.672,P=0.01 ).The plasma sHLA-G levels in the infectious mononucleosis children were dramatically higher than that in normal controls,and a significant difference was observed between the two groups( Z=-9.472,P＜0.01 ).Among the infectious mononucleosis children,levels of sHLA-G was find a significant difference between the three genotypes of HLA-G 14 bp insertion/deletion polymorphism( H=6.09,P =0.048 ),and the level of s HLA-G with 14 bp+/+ genotype was markedly lower than that of the two other genotypes (Z=-2.376,P=0.01 8).Conclusion There was a relationship between the HLA-G 14 bp insertion/deletion polymorphism and the susceptibility to the infectious mononucleosis for children.Children who carried the 14 bp-/- genotype or deleted the 14 bp allele may have a significantly increased risk of the infection of EBV.The plasma sHLA-G might be considered as an index for auxiliary diagnosis infectious mononucleosis.

Objective To establish a method for detection of gene mutations in β-thalassemia by high-resolution melting (HRM) and study its preliminary clinical application.Methods Two common mutations [ IVS-2-654 ( C ＞ T ) and -28 ( A ＞ G ) ]of β-thalassemia in Wenzhou city population were selected.The plasmid DNA fragments of these mutations were constructed by TA clone technology as PCR templates or genotyping controls.A method for detection of β-thalassemia gene mutations based on HRM analysis was established and its specificity,sensitivity and repeatability were methodologically evaluated.One hundred and seventeen patients with clinically suspected β-thalassemia from Second Affiliated Hospital and Yu ying Children's Hospital of Wenzhou Medical College were enrolled into this study.The genomic DNA was extracted from whole blood cells and detected by HRM method.The results were compared with the direct sequencing data.Results HRM method could detect the mutations [ IVS-2-654( C ＞ T) and -28 ( A ＞ G ) ]of β-thalassemia and the results did not show any non-specific amplified fragments.All within-run and between-run coefficients of variation for different DNA types' Tm were smaller than 0.1％.And minimum 103 copies of DNA of each assay and 10％ mutation could be determined by this method.One hundred and seventeen patients with clinically suspected β-thalassemia were detected with HRM and all the results were in accordance with direct DNA sequencing.There were 45 IVS-2-654 ( C ＞ T)heterozygous mutation and 9-28 ( A ＞ G)heterozygous mutation and none homozygous mutation.Conclusion The method of rapid identification of β-thalassemia gene mutations based on HRM analysis is successfully established,which is a convenient,rapid,specific,sensitive and accurate technique for screening gene mutations in β-thalassemia as well as a general technical platform to identify other gene mutations.

Objective To explore the clinical application value of neutrophil gelatinase-associated lipocalin(NGAL)which were tested by immunity transmission turbidity in early kidney injury after elective percutaneous coronary intervention.Methods A case-control study was conducted.All 201 stable angina pectoris and acute coronary syndrome patients undergone percutaneous coronary intervention in TEDA International Cardiovascular Hospital,during April to August 2013,were enrolled in this study.Before and 2 h,4 h,8 h,24 h,48 h after the operation,the plasma creatinine of the patient samples were tested by enzymic method.Before and 2 h,4 h,8 h,24 h after the operation,the plasma NGAL was tested by immunity transmission turbidity method.Before and 8 h,24 h after the operation,the urinary NGAL was tested by immunoturdimetric method.The data were compared between contrast induced nephrpathy (CIN) and non-CIN groups.For normal distribution of quantitative data,t test were used and for non-normal distribution of quantitative data,nonparametric rank and inspection were used.Results CIN occurred in 8 of 201 enrolled patients,the incidence was 3.98％.Receiver operating characteristic curve (ROC) analysis confirmed the diagnostic accuracy of the plasma NGAL in CIN,and the area under the curve(AUC) of 2 h plasma NGAL was 0.928,95％ CI 0.800-0.985,with the cut-off value NGAL as 109 ng/ml,the diagnostic sensitivity and specificity for CIN were 87.5％ and 100％ ;the AUC of 8 h plasma NGAL was 0.945,95％ CI 0.824-0.992,with the cut-off value NGAL as 96 ng/ml,the diagnostic sensitivity and specificity for CIN were 87.5％ and 87.5％ ;the AUC of 8 h urinary NGAL was 0.969,95％ CI 0.859-0.999,with the cutoff value NGAL as 91 ng/ml,the diagnostic sensitivity and specificity for CIN were 87.5％ and 100％.Conclusions The change of plasma and urinary NGAL is earlier to that of serum creatinine for the early diagnosis of CIN.It can be used as the predictor of early renal damage after elective coronary artery interventional.

ObjectiveTo compare the effect of acupuncture and speech rehabilitation on motor aphasia after stroke with speech rehabilitation alone. Methods49 patients with aphasia were divided into treatment group (27 cases) and control group (22 cases) randomly. The cases in treatment group were treated with acupuncture and speech rehabilitation. The cases in control group were treated with speech rehabilitation alone. They were assessed before and 1 month, 3 months after treatment. ResultsThere was no significant difference between treatment group and control group 1 month after treatment (P>0.05), but was significant in auditory comprehension, speaking, reading, dictation and description (P<0.05) 3 months after treatment. ConclusionThe acupuncture can facilitate the recovery of motor aphas after stroke.

OBJECTIVETo label embryonic stem (ES) cells with enhanced green fluorescent protein (EGFP) on the hypoxanthineguanine phosphoribosyl transferase (HPRT) gene locus for the first time to provide a convenient and efficient way for cell tracking and manipulation in the studies of transplantation and stem cell therapy.

METHODSHomologous fragments were obtained by polymerase chain reaction (PCR), from which the gene targeting vector pHPRT-EGFP was constructed. The linearized vector was introduced into ES cells by electroporation. The G418(r)6TG(r) cell clones were obtained after selection with G418 and 6TG media. The integration patterns of these resistant cell clones were identified with Southern blotting.

RESULTSEGFP expressing ES cells on the locus of HPRT were successfully generated. They have normal properties, such as karyotype, viability and differentiation ability. The green fluorescence of EGFP expressing cells was maintained in propagation of the ES cells for more than 30 passages and in differentiated cells. Cultured in suspension, the "green" ES cells aggregated and formed embryoid bodies, retaining the green fluorescence at varying developmental stages. The "green" embryoid bodies could expand and differentiate into various types of cells, exhibiting ubiquitous green fluorescence.

CONCLUSIONSThis generation of "green" targeted ES cells is described in an efficient protocol for obtaining the homologous fragments by PCR. Introducing the marker gene in the genome of ES cells, we should be able to manipulate them in vitro and use them as vehicles in cell-replacement therapy as well as for other biomedical and research purposes.

Animals ; Cells, Cultured ; Chromosome Mapping ; Embryo, Mammalian ; cytology ; Green Fluorescent Proteins ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Luminescent Proteins ; metabolism ; Mice ; Recombination, Genetic ; Stem Cells ; metabolism ; Transgenes