Objective To investigate the protective effect of electroacupuncture pretreatment on myocardial ischemia/reperfusion (I/R) injury and its influence on high mobility group box 1 (HMGB1) expression in rats. Methods Sixty Wistar rats were randomly divided into sham operation group, myocardial I/R model group and electroacupuncture pretreatment group by random number table (each n = 20). Myocardial I/R injury model was reproduced by ligating the left ventricular branch coronary artery at about 0.5 cm below the atrial appendage lower margin for 10 minutes to occlude the blood flow, then the ligature was relaxed for 1 hour reperfusion; in electroacupuncture pretreatment group, 7 days before I/R, the electroacupuncture at Neiguan acupoint was applied once daily for 20 minutes till the 7th day when I/R was established. Under light microscope, the pathological changes of myocardial specimen stained by hematoxylin-eosine (HE) method were observed. The myocardial histopathological integral was detected by semi quantitative integral method, and the changes of histological scores in three groups were investigated. The levels of plasma HMGB1, tumor necrosis factor-α (TNF-α), cardiac troponin T (cTnT) were detected by enzyme-labeled immunosorbent assay (ELISA). The expressions of HMGB1, monocyte chemotactic protein-1 (MCP-1), TNF-αmRNA and protein in myocardium were detected by reverse transcription-polymerase chain reaction (PT-PCR) and Western Blot. Results Under light microscope, the myocardial tissue in myocardial I/R model group showed partial fracture of myocardial fibers, large patches of myocardial cell necrosis, hazy boundary, cellular condensation, rupture and dissolution or even disappearance, interstitial edema with a lot of inflammatory cell infiltration; the above myocardial tissue injury in electroacupuncture pretreatment group was significantly milder than that in myocardial I/R model group. Compared with sham operation group, in myocardial I/R model group the HMGB1, TNF-α, cTnT contents and histological score were significantly increased [HMGB1 (μg/L):9.64±1.16 vs. 2.15±0.31, TNF-α(μg/L):91±22 vs. 19±5, cTnT (μg/L):1.50±0.35 vs. 0.07±0.03, histological score:2.5±0.3 vs. 0.0±0.0, all P<0.01], HMGB1, MCP-1, TNF-α mRNA and protein expressions were increased obviously (HMGB1 mRNA: 1.42±0.16 vs. 0.02±0.00, MCP-1 mRNA:0.46±0.06 vs. 0.01±0.00, TNF-αmRNA:0.75±0.04 vs. 0.03±0.00;HMGB1 protein:1.08±0.01 vs. 0.20±0.01, MCP-1 protein:0.92±0.03 vs. 0.40±0.01, TNF-αprotein:1.10±0.02 vs. 0.35±0.01, P<0.05 or P<0.01);compared with myocardial I/R model group, in electroacupuncture pretreatment group, HMGB1 (6.58±0.73), TNF-α (63±19), cTnT (1.15±0.31) levels were significantly decreased (all P < 0.01), HMGB1, MCP-1, TNF-αmRNA and protein expressions were markedly reduced (mRNA expression was 0.74±0.12, 0.18±0.02, 0.10±0.03, and protein expression was 0.40±0.01, 0.36±0.02, 0.50±0.02, respectively all P<0.05), and histological score (1.2±1.0) was remarkably lowered (P < 0.01). Conclusion Electroacupuncture pretreatment may reduce the myocardial I/R injury in rats, and the mechanism may be related to the amelioration of inflammatory response mediated by HMGB1 at late stage.

Objective To analyze the genotypes of adenovirus(Ad)strains isolated from young children with acute respiratory infection syndrome during 2004-2006.Methods Seven human adenovirus strains isolated during 2004 and 2006 were detected with tissue culture,PCR with adenovims specific-primers and sequencing for hexon and fiber genes.Results The results showed that all 7 adenovirus strain belonged to adenovirus genera B.3 of the 7 isolates were Ad3,BJ060605-1,BJ060316-and BJ040228 had high homology of nucleic acid.98.9%-100%.Three of 7 were Ad7,the homology of nucleic acid of BJ050520.BJ060421-4 and BJ040226 were 97.6%-100%.Strain SX060418-1 was 100% homologic with Ad11 strain L08232(GenBank).Conclusion Little variation of fiber gene was found for 2004 to 2006 isolares and the typing with fiber sequencing is similar with that of hexon gene of adenovirus isolates.

Objective To investigate the clinical value of acoustic radiation force impulse (ARFI) imaging in the evaluation of liver functional reserve in patients with hepatic focal lesions. Methods Eighty-six patients with hepatic focal lesions in General Hospital of the People′s Liberation Army from September 2011 to August 2012 were studied. According to Child-Pugh score, 54 of them were classified as grade A, 18 patients were classiifed as grade B, and 14 patients were classiifed as grade C. Seventy patients underwent surgery and 16 patients underwent non-surgical treatment. Shear wave velocity (SWV) values were measured using ARFI technique, and indocyanine green (ICG) clearance tests were also performed to achieve ICG clearance rate constant (ICGK) and ICG retention rate at 15 minutes (ICG15). Correlation between SWV and ICGK, SWV and ICG15 were studied using Pearson correlation, and correlation between SWV and Child-Pugh score were analyzed using Spearman rank correlation. Difference between SWV values of patients underwent surgery and those of patients underwent non-surgical treatment was compared using t-test for independent samples. Analyses of receiver operating characteristic (ROC) curve were performed to investigate the value of ARFI in the evaluation of surgical feasibility for patients with hepatic focal lesion, with the clinical decisions serving as the golden standard. Results There was correlations between SWV and ICGR15, SWV and ICGK, and SWV and Child-Pugh scores (r=0.764,P <0.001;r=-0.686,P=0.000;r=0.864,P=0.000). The SWV of patients who underwent surgery was (2.46±0.45) m/s, and the SWV of patients who underwent non-surgical treatment was (1.54±0.36) m/s, with a signiifcant difference (t=-0.80, P=0.000). The ROC curve analysis demonstrated a highest Youden′s index of 0.775 when the cut-off was 2.06m/s. The corresponding sensitivity was 87.5% and the specificity was 90.0% for ARFI in the diagnosis liver functional reserve deficiency. Conclusion ARFI can effectively evaluate the liver functional reserve in patients with hepatic focal lesions and thus could be beneifcial in the decision of clinical treatment.

Objective To study the clinical value of acoustic radiation force impulse imaging (ARFI) in patients with ischemic type biliary lesion (ITBL) after orthotopic liver transplantation. Methods Between July 2012 to January 2013, forty-one patients in General Hospital of the People′s Liberation Army were enrolled in this study who were admitted for post-transplantation follow-up. Among them, 15 patients were diagnosed as ITBL by ultrasonography. Besides conventional ultrasonography, ARFI was used to detect the shear wave velocity (SWV) of liver tissue at depth of 4 cm and 5 cm respectively. Simultaneously liver function test was performed. Independent-samples t test was performed to compare the difference of SWV between ITBL and non-ITBL group at the same depth. Paired-sample t test was performed to compare the difference of SWV for the same ITBL patient. Pearson correlation analysis was used in analyzing the relation between SWV and liver function. Results The average SWV in depth of 4 cm was (1.561±0.425) m/s and (1.121±0.160) m/s in ITBL and non-ITBL group, respectively. Signiifcant differences were found among the ITBL and non-ITBL groups (t=-3.173, P=0.01). The average SWV in depth of 5 cm was (1.608±0.545) m/s and (1.175±0.173) m/s in ITBL and non-ITBL group, respectively. Signiifcant differences were found among the ITBL and non-ITBL groups (t=-2.454, P=0.034). There was no signiifcant difference between SWV measurements at different depth for the same ITBL patient. For all patients, SWV at different depths were both strongly correlated with alkaline phosphatase (r=0.656, 0.667, respectively;both P=0.000) andγ-glutamyl transpeptidase (r=0.482, P=0.007;r=0.508, P=0.004). Conclusion The liver stiffness measurement is valuable for the clinical evaluation of post-transplantation ITBL.

BACKGROUND:Human amniotic membrane-derived mesenchymal stem cells (AMSCs) are considered to be one kind of adult stem cells that can be easily obtained in large quantities without using an invasive method. Because of their low immunogenicity, anti-inflammatory properties, multipotency of differentiation and without ethical issue, human amniotic membrane-derived mesenchymal stem cells have been proposed as a good candidate to be used in celltherapy and regenerative medicine. However, the biological properties and the differentiation capacity of human amniotic membrane-derived mesenchymal stem cells are stil poorly characterized. OBJECTIVE:To establish a practical method for isolation and purification of human amniotic membrane-derived mesenchymal stem cells, and to study the biological characteristics and dopaminergic neural-like celldifferentiation potential of the human amniotic membrane-derived mesenchymal stem cells. METHODS:Human amniotic membrane-derived mesenchymal stem cells were disassociated and isolated from the amniotic membrane by trypsin and col agenase based enzymic digestion, and purified by percol mediated density gradient centrifugation. Expressions of surface antigens and transcription factors of the human amniotic membrane-derived mesenchymal stem cells were determined by flow cytometry and western blot assays. Based on the osteogenic and adipogenic induction, the multipotent differentiation capability of human amniotic membrane-derived mesenchymal stem cells was determined. Induction of neural celldifferentiation of human amniotic membrane-derived mesenchymal stem cells was conducted in Neurabasal conditioning medium with ATRA supplement. Neural cellassociated bio-markers were determined by immunofluoresence staining and confocal microscope. RESULTS AND CONCLUSION:In this study, we performed a practical method to isolate and purify human amniotic membrane-derived mesenchymal stem cells and amniotic epithelial cells simultaneously, with high cells yield. We demonstrated a group of constitutive expressions of neural antigens and embryonic associated transcription factor proteins (OCT-4, SOX-2 and KLF4) in fresh isolated human amniotic membrane-derived mesenchymal stem cells as wel as in human amniotic membrane-derived mesenchymal stem cells after in vitro passage, which suggested that the human amniotic membrane-derived mesenchymal stem cells not only possessed intrinsic tendency to neural celldifferentiation, but also maintained their stem cellcharacteristics after in vitro passage. We stimulated the human amniotic membrane-derived mesenchymal stem cells in the neurobasal-A and B27 based conditioning medium to induce neural celldifferentiation. The induced human amniotic membrane-derived mesenchymal stem cells displayed an up-regulation of expression in panel of neural and dopaminergic associate molecules (β-tubulin III, neuron-specific nuclear protein, tyrosine hydroxylase, glial fibril ary acidic protein, myelin basic protein and nestin) by flow cytometry and immunofluorescence staining, which demonstrated the multipotent differentiation capability and dopaminergic neuron-like differentiation potential of the human amniotic membrane-derived mesenchymal stem cells.

Objective To analyze the prevalence,risk factors of kidney disease in type 2 diabetic patients with KDIGO classification of chronic kidney disease,also to study cardiovascular and cerebrovascular diseases and death in these patients,so as to investigate the significance of the KDIGO classification system.Methods One thousand six hundred and forty-five type 2 diabetic patients who were in hospitalization from June 2008 to December 2012 were grouped according to the KDIGO classification of chronic kidney disease and the incidence of vascular disease was analyzed based on the classification.Clinical features were compared between patients with or without kidney disease.The risk factors of kidney disease and the death of diabetic patients were also investigated.Results There were 915 male and 730 female,aged a median (57.86±12.54) years with (6.35±6.30) years duration of diabetes mellitus among the 1645 cases,and 37.2％ of patients had concomitant kidney disease.According to the classi fi cation of CKD,patients in CKD group 3a,group 3b and CKD group 4-5 accounted for 5.7％,3.5％ and 7.6％,while 33.4％ of patients had proteinuria,among which 19.5％ with microalbuminuria,13.5％ with macroalbuminuria.On complications,patients with hypertension accounted for 49.5％,hyperlipidemia 67.7％,diabetic retinopathy 27.4％,cardiovascular and cerebrovascular diseases 18.5％ (coronary artery disease 16.5％,cerebrovascular diseases 8.8％).Statistical difference was detected in the incidence of diabetic retinopathy,coronary artery disease and cerebrovascular diseases between CKD group 3a and 3b (P ＜ 0.05).The duration of diabetes,concomitant hypertention especially with elevated systolic blood pressure,diabetic retinopathy and hyperuricemia were the independent risk factors for type 2 diabetic patients with kidney disease.Age,Scr,complicating cardiovascular and cerebrovascular diseases and advanced CKD stage were the independent risk factors for the death of type 2 diabetic patients with kidney disease.Conclusion KDIGO classification of chronic kidney disease enables better staging of kidney diseases in diabetic patients for management and prognosis.Diabetic patients have a higher prevalence of renal diseases and cardiovascular and cerebrovascular events than the general population.Early control of factors such as blood pressure and serum uric acid can delay the progression of kidney disease,and the predictive role of diabetic retinopathy should be emphasized.

Objective To enhance the test efficiency and management level of medical record quality by designing and using med-ical record quality management system based on new electronic medical record .Methods System design and development were based on JAVA ,consisting of data exchange platform and medical record quality management system .The platform used C/S archi-tecture and the main program used B/S architecture .Achieved the integration with new electronic medical record and clinical infor-mation system through data exchange with business database .Results The supervision model and business workflow of medical re-cord quality were changed ,and the process and terminal medical record quality were promoted .Conclusion Design and application of the system can provides strong support for medical record quality management ,supplys effective supports for supervision of med-ical record quality ,and plays positive role in promoting medical record quality ,clinical work efficiency ,and medical quality .

Object To develop an immunoassay method of species-specific protein for Radix Glycyrrhizae (RG) identification. Methods The crude drug of RG as a model was studied in this paper,the antigen of RG protein (RGP) was screened by Western-blot analysis using anti-RG serum. After species-specific protein,RGP was purified,and rabbit antiserum against RGP was prepared,the rabbit IgG of anti-RGP was isolated and biotinylated,then the sandwich enzyme-linked immuosorbent assay (ELISA) was developed. Results The method was only specific to RG,which was harvested from various areas or species,and showed excellent sensitivity and reproducibility. Conclusion The result suggests that the immunoassay method using specific antigen of RGP as a detection objective be a new potential for the unequivocal identification and quality control of RG.

Objective To evaluate the value of supersonic shear wave elastrography(SWE) in the diagnosis of benign and malignant breast lesions.Methods SWE was performed on 134 breast lesions of 123 female patients and 74 normal breast glands to determine the values of overall elastic modulus and dispersion,and ROC curves were used to assess the diagnosis boundary value of two kinds of data respectively.Results According to the diagnostic gold standard-histopathology,the values of overall elastic modulus and dispersion were statistically significant when comparing benign lesions and malignant lesions with normal glands.ROC curve displayed that,when the Youden index reached to maximum,the diagnosis boundary value of overall elastic modulus was 41.01 kPa,the sensitivity and specificity of the value were 68.5％ and 83.8％ respectively; the diagnosis boundary value of dispersion was 12.25 kPa,the sensitivity and specificity of the value were 87.0％ and 88.7％ respectively.Conclusions The values of overall elastic modulus and dispersion for solid breast lesions can be used to reflect the elastic characteristics of lesions quantificationally,which is useful to diagnose benign and malignant breast lesions in clinical.

Objective To discuss the timing of extubation in patients of congenital heart disease (CHD)with severe pulmonary hypertension (PH) undergoing connective surgery with cardiopulmonary bypass(CPB).Methods A retrospective study of 40 patients of CHD with severe PH Was completed.According to whether the duration of mechanical ventilation (MV)>24 hours,patients were divided into two groups,early extubafion group(19 patients) and late extubation group(21 patients).Perioperative variables that might influence duration of MV were compared between the two groups.Multivariate statistical analysis with Logistic regression was used for these patients to analyze the perioperative variables to determine risk factors for prolonged MV (MV≥72 hours).Results Between the two groups,preoperative NYHA class,Pp/Ps at the time of coming off bypass,CPB duration,and CPB temperature were significantly different (P<0.05 or<0.01).Anofic spell (OR=0.022,95%CI0.001-0.580)and severe hypoxemia(OR=0.031,95%CI0.002-0.568)in the early postoperative period were the risk factors for prolonged MV.Conclusions The timing of extubation in these patients should be determined individually.Late extubation may fit those with advanced NYHA class,high Pp/Ps at the time of coming off bypass,prolonged CPB duration,or hypothermic cardiography and pulmonary artery catheter monitoring help to guide treatment for pwlonged MV.For else patients who with postoperative stable cardiopulmonary function,early extubation maybe feasible.