ObjectiveTo investigate the role of the anti-podocyte antibody in the pathogenesis of membranous nephropathy(MN).Methods Thirty-six frozen section slice from biopsies of normal kidney were randomly divided into 3 groups:A,B and C groups(n=12,respectively),then,serum of MN and IgA nephropothy patients,and normal saline were added by drops to frozen section respectively,then the changes were observed by fluorescence microscope.Twenty-one male New Zealand white rabbits were feed with standard diet for 1 week,then were randomly divided into 3 groups:D,E,F groups (n=7,respectively),and injected with the serum of MN and IgA nephropathy,and normal saline respectively through ear-border vein.24-h urinary protein, serumalbumin, serumcreatinineweredetected. Atweek8afterinjection, the histopathological changes in kidney tissue of rabbits were observed by light microscope,electron microscope and immunofluorescence staining. Results Immunofluorescenee staining displayed IgG deposition in glomerular podocytes in group A,and there was no positive expression in other two groups.The quantitative measurement of 24-h urinary protein increased significantly after injection of MN serum (P＜0.01),meanwhile the serum albumin markedly decreased (P＜0.01).The inmnunofluorescence staining showed that IgG deposited in the glomerulus capillaries wall,which presented spherical particles.Light microscope revealed the thickening of GBM without nailing process formation.Electron microscope showed the podocyte foot processes,subepithelial dense deposits.Such pathological changes were not found in E group and F group.Conclusion There is anti-podocyte antibody in blood of MN patients,which may play a role in the pathogenesis of MN.

Objective:To establish an HPLC method for the determination of gastrodin in Fufang Zhenma capsules. Methods:A YMC-Pack Pro C18(250 mm ×4.6 mm,5μm)column was used. The mobile phase consisted of acetonitrile-water (2 ∶98). The detec-tion wavelength was 220 nm. The flow rate was 1. 0 ml·min-1 ,the column temperatime was 20℃. the sample size was 25μl. Results:Gastrodin showed a good linear relationship within the range of 3.38-432.80 μg·ml-1(r =0.999 9). The average recovery was 97. 14% (RSD=2. 67%)(n=6). Conclusion:The method is feasible with good specificity and reproducibility, and can be used for the determination of gastrodin in Fufang Zhenma capsules.

ObjectiveTo investigate the current situation of immunophenotyping of acute leukemia by flow cytometry in China (except Taiwan, Hongkong and Macau), and to provide theoretical evidence for the formulation of guidelines of immunophenotyping of leukemia by flow cytometry.MethodsThe Chinese Hospital Knowledge Database(CHKD) was used to retrieve the1994-2009 published papers on immunophenotyping of acute leukemia by flow cytometry.All of 380 retrieved papers were analyzed by the GraphPad Prism 4. ResultsThe number of published papers in China increased yearly and changed gradually from large cities to small and medium-sized cities in local distribution. The most often reported area was Beijing, followed by Jiangsu and Guangdong. In specimen processing, the ratios of using mononuclear cell labling method and whole blood labling-lysing-washing method were 34.2 ％ (113/330) and 65.8 ％ (217/330) respectively. The ratios of using single color, two color, three color and four color labling method were 15.4 ％ (53/344), 13.7 ％ (47/344), 54.1％ (186/344) and 16.3 ％ (56/344), respectively. The number of fluorescent antibodies was used from less than 10 to more than 20.Most of them were between 11 to 16,which was 48.6 ％ (167/344) of total number of published papers. In data acquisition and analysis, the ratios of using FSC/SSC gating and CD45/SSC gating were 34.2 ％ (113/330) and 65.8 ％ (217/330), respectively. There were some differences in the positive criteria of lymphatic and myeloid and cytoplasm antigens.The main positive criteria of lymphatic, myeloid antigen was ≥20 ％, which was 64.8 ％ (223/344) and 95.4 ％ (328/344) . The main positive criteria of cytoplasm antigen was ≥ 10 ％, which accounted for 61.2 ％ (156/255). Conclusion A common view has been reached in specimen processing,gating method,data acquisition and analysis in immunophenotyping of acute leukemia by flow cytometry. But there were still some difference in the number of fluorescent antibodies, panels and evaluation of positive criteria of antigens.

Objective To study the effects of high matrix metalloproteinase 9 (MMP 9 ) on the biological behaviors of fibroblasts,in order to clarify the possible mechanisms in the wound healing of diabetic foot.Methods Establish the cell model of skin fibroblast with high expression of MMP 9 by high glucose (22.0 mmol/L) and high homocysteine (100 μmol/L) co-culture.Control group was incubated with normal glucose (5.5 mmol/L).Realtime PCR,ELISA and gelatin zymography were used to detect the MMP 9 mRNA,protein expression and activity of MMP 9.Flow cytometry,CCK-8,ELISA assay,scratch test and transwell were used to detect cell proliferation,viability,collagen (hydroxyproline) secretion,horizontal migration and vertical migration of cells.Results Data were expressed as (x- ± s).Differences were considered significant if their P value was ＜ 0.05.Results The expression of MMP 9 mRNA,protein levels and the activity of MMP 9 in high MMP 9 group were 6.05 folds,4.12 folds and 1.58 folds higher than those in control group (P ＜ 0.01,respectively).The proportion of S phase cells,proliferation index,cell viability,collagen (hydroxyproline) secretion,6 h horizontal migration rate and the number of vertical migration cells in high MMP9 group decreased by 29.8 ％,18.1％,23.3 ％,68.7 ％,45.0 ％ and 21.4 ％ than those in control group (P ＜ 0.01,respectively ). Conclusions Fibroblast with high expression of MMP 9 have decreased proliferation,activity,collagen secretion and reduced migration,which suggests that MMP 9 may inhibit the biological behaviors of fibroblasts.

Objective The study aimed to analyze the clinical features and the causes of hospital death among the 5720 acute myocardial infarction(AMI)patients from Cardiology Center,Beijing Chao-Yang Hospital during the last 8 years. Methods A total of 5720 AMI patients received treatment in the Cardiology Center from January 1st ,2002 to December 31th ,2009 were retrospectively reviewed. All patients were classified according to age into 3 groups of ≤45,46 -75,and ＞ 75 years old. The morbidity,cause of death ,whether they had the PCI therapy,mortality after PCI and the impact of gender on the cause of death were observed respectively. Results The morbidity rate of male was significantly higher than female in all three groups,and the study also found that the morbidity rate of female was significantly higher in the group of ＞ 75 years old,which however was still lower than that of male. The AMI patients were more likely to accept PCI therapy,which could significantly reduce the mortality rate. The top 3 causes of death included acute heart failure(AHF),cardiogenic shock(CGS)and AHF combined with CGS. In addition,AHF caused significantly more death in female and older(＞ 75 years old)patients. Conclusions The morbidity rate of AMI patients in Beijing Chao-Yang Hospital increased year by year. And PCI therapy could reduce the mortality rate of all groups. Revascularization treatment seems to be feasible and safe for the patients older than 75 years old.

Glioma ; pathology ; Humans ; Neoplastic Stem Cells ; pathology

Objective To explore the molecular characteristics of community-associated Methicillin-resistant Staphylococcus aureus (CA-MRSA) from children with skin and soft tissue infection (SSTIs) in China.Methods CA-MRSA was collected from the outpatients in Department of Dermatology in 13 Children's Hospitals of China.Genotypic characteristics of CA-MRSA isolates were tested by adopting mnltilocus sequence typing (MLST),staphylococcal protein A (spa) typing and Staphyloccoccal cassette chromosome mec (SCCmec) typing.And the presence of Panton-Valentine leukocidin genes(pvl) was determined also.Results Overall,1 705 strains of Staphylococcus aureus were isolated from SSTIs children,and CA-MRSA accounted for 2.6％ (44/1 705 strains).Two types of SCCmec were detected in all the CA-MRSA strains,and the main types were SCCmec Ⅳ and SCCmec Ⅴ,accounting for 45.5 ％ (20/44 strains) and 54.5 ％ (24/44 strains),respectively.Thirteen MLST types (STs) and 15 spa types were detected among CA-MRSA.The most prevalent MLST was ST121 (18/44 strains,40.9％),followed by ST59 (9/44 strains,20.5％).Additionally,t437 was predominant,accounting for 40.9％ (18/44 strains).ST121 strain had 8 spa types,and t2086 was the most common type (6/18 strains,33.3％);while ST59 had only 1 spa type,t437.No ST121,ST59 and t437 strains were found in Central and Eastern region of China.Only 4 (9.1％) of the 44 CA-MRSA strains harbored pvl genes which were all from Southern region.Conclusions The most common clone of SSTIs CA-MRSA in children is MRSA-ST121-SCCmec Ⅳ/Ⅴ,as the molecular epidemiology of CA-MRSA strain has changed,ST121 has replaced ST59 to become the main epidemic strains.

Objectives:This paper at measuring the inequity and its influencing factors of medical care utiliza-tion of elderly aged above 60 ( inclusive) . Methods:data comes from 2013 China Health and Retirement Longitudinal Study ( CHARLS) where the population aged 60 and above was selected as the research object. Concentration index ( CI) and its decomposition or centralized curve was used to measure the inequity of medical care services utilization of the elderly, and then the influencing factors of inequity were analyzed by means of the centralized index. Results:The concentration index for outpatient and inpatient service utilization for the elderly was 0 . 0619 and 0 . 1050 , re-spectively, and the concentration curves were below the absolute fair line. The top 2 factors that showed positive con-tribution to the outpatient service utilization included annual per capita consumption expenditure and the pension a-mount. The top 2 factors that showed negative contribution and larger contribution rate to the outpatient service utiliza-tion included New Rural Cooperative Medical Insurance (NRCMI), and Physical Ability in Daily Life (PADL). The top 2 factors that positively and highly contributed to the inpatient service utilization included the household per capita consumption expenditure and the Urban Employees' Basic Medical Insurance (UEBMI). The top 2 factors that nega-tively contributed to the inpatient service utilization included the New Rural Cooperative Medical Insurance ( NRCMI) and the Physical Ability in Daily Life ( PADL) . The horizontal inequity of outpatient and inpatient service utilization was 0. 0739 and 0. 1339, respectively, indicating that there was unfairness in the use of outpatient and inpatient services among elderly. Conclusion:There is inequity of medical care service utilization among the elderly in China. The economic status contributes the largest part of inequity, meaning that it is unfair to the first contribution factor;while the Needs-based fac-tors and New Rural Cooperative Medical Insurance (NRCMI) showed an inequity, narrowing the unfair gap.

Objective To establish the dissolution curves test method of Ribavirin capsules, and investigate dissolution behavior of domestic Ribavirin capsules.To provide experimental basis for generic drugs quality consistency evaluation.Methods According to the first dissolution method (basket method)stated in appendix Ⅹof Chinese Pharmacopeia(2010 edition),the rotation speed was 50 r/min with dissolution medium volume of 900 mL.The dissolution profiles of Ribavirin capsules in four different mediums( pH 1.2 hydrochloric acid solution,pH 4.5 acetic buffer,pH 6.8 phosphate buffer and water) were determined by HPLC.The determination was performed on C18 column with mobile phase consisted of 4 g/L sodium dihydrogen phosphate solution(pH adjusted to 5.0 ±0.05 using 5% sodium hydroxide solution)-acetonitrile(98:2)at the flow rate of 1.0 mL/min.The detection wavelength was 225 nm,and sample size was 10μL.Results The linear range of ribavirin was 2.5-200μg/mL(r=1).RSD of precision and stability tests were lower than 0.5%.The average recoveries were 101.3%, 100.7%, 100.2%, 100.4% in four mediums.Dissolution behavior of capsules can be more consistent and rapid dissolution in pH4.5 and pH6.8 mediums.But they were quite different in pH1.2 and water mediums, and some of their average dissolution at 15 min could not reach 85%.Conclusion This method is accurate and reliable.There is a difference between domestic Ribavirin capsules dissolution behavior, and the formulation processes have room for improvement.

Objective To investigate the roles of phosphatidylinositol 3 kinase (PI3K) and phosphorylated Akt (P-Akt) in the pathogenesis of cervical squamous cell carcinoma and condyloma acuminatum.Methods Immunohistochemistry and Western blot were used to detect the expressions of PI3K and P-Akt in tissue specimens from the lesions of 30 cases of cervical squamous cell carcinoma,30 cases of condyloma acuminatum and the prepuce of 15 normal human controls.The average optical density and gray scale values were calculated and analyzed by t test and F test respectively.Results The expressions of PI3K and P-Akt were observed in only the basal layer of the epidermis of control specimens,but in the whole epidermis of condyloma acuminatum tissue specimens.Cervical squamous cell carcinoma tissue specimens displayed a stronger expression of PI3K and P-Akt compared with the control and condyloma acuminatum tissue specimens.As immunohistochemistry revealed,the average absorbance value for PI3K and P-Akt was 0.28 ±0.05 and 0.20 ± 0.07 respectively in cervical squamous cell carcinoma tissue specimens,0.22 ± 0.04 and 0.17 ± 0.03 respectively in condyloma acuminatum tissue specimens,and 0.16 ± 0.04 and 0.10 ± 0.02 respectively in the control tissue specimens; significant differences were observed in the expressions of PI3K and P-Akt among the three groups of tissue specimens (F =44.87,20.64,respectively,both P ＜ 0.01 ).The results of Western blot were consistent with those of immunohistochemistry,and there was a significant difference in the gray scale value for PI3K and P-Akt between cervical squamous cell carcinoma,condyloma acuminatum and control tissue specimens (3.48 ± 0.48 vs.1.99 ± 0.11 vs.1.00 ± 0.03,F=354.83,P＜ 0.01; 3.33 ± 0.26 vs.1.96 ± 0.11 vs.1.00 ± 0.03,F=302.33,P＜ 0.01 ).Conclusions The PI3K/Akt signaling pathway is abnormally activated in condyloma acuminatum and cervical squamous cell carcinoma,and human papilloma virus may cause the abnormal proliferation of infected epithelium likely by affecting the upregnlated expression of PI3K/P-Akt.