Objective To observe the changes of foot processes,expression and distribution of transient receptor potential cation channel 6 (TRPC6) in podocytes by puromycin aminonucleoside (PAN) and dexamethasone (DEX) intervention,then to investigate the function of TRPC6 in podocytes and its relation to proteinuria in kidney diseases.Methods Podocytes cultured in vitro were divided into three group:control group,PAN stimulation group and DEX intervention group.Mouse podocyte cell line (MPC5) were cultured in 0.02％ dimethyl sulfoxide (DMSO) in control group,subjected to PAN (50 μg/ml) treatment alone or with DEX (1 μmol/L) in other two groups for 8 h,24 h,48 h.The podocyte morphology was observed and took pictures by phase-contrast microscope,then the differences of morphology and areas were analyzed.The distribution,mRNA expression and protein expression of TRPC6 were detected by indirect immunocyto-fluorescence,real-time quantitative PCR and Western blotting,respectively.Results The well-developed podocyte arborization and interconnection was formed in control group,but PAN led to significant shrinkage of podocytes (P ＜ 0.05),together with podocyte foot process retraction,effacement and loss of cell contact.DEX significantly prevented the shrinkage and apoptosis of podocytes.The apoptosis rate was significantly increased after PAN stimulated 48 h (P ＜ 0.05).Real-time quantitative PCR and Western blotting found TRPC6 mRNA and protein expression were prone to increase in PAN group compared with control group (P ＜ 0.05).The distribution of TRPC6 becamed abnormal in PAN group.DEX decreased TRPC6 mRNA and protein expression at 48 h compared with PAN group (P ＜ 0.05).The abnormal distribution of TRPC6 was also alleviated by the protection of DEX.Conclusion DEX exerts a direct action to podocyte which retains the integrity of slit diaphragm against podocyte injury,and alleviates proteinuria via stabilizing mRNA,protein expression and distribution of TRPC6.

Objective To observe the expression and distribution of α-actin-4 mRNA through puromycin aminonueleoside(PAN) injury podocyte,and discuss the relation between α-actin-4 and podocyte damage.Methods Podocytes were cultured in vitro,and 2 groups were set up:control group and PAN stimulation group.The control group was cultured with concentration of 100 mL/L FBS RPMI 1640 nutrient solution culture,while the PAN group was cultivated to PAN(50 mg/L) treatment,and cell morphology,extraction of total RNA of α-actin-4 were observed in 8 h,24 h and 48 h.The podocyte morphology was observed and pictures were taken through phase-contrast microscope,then the differences of morphology and areas between the 2 groups were analyzed.The distribution and mRNA expression of α-actin-4 were detected by indirect immunocyto-fluorescence and real-time quantitative PCR,respectively.Results The well-developed podocyte arborization was formed after the in vitro induction,and the PAN treatment led to the podocyte foot process retraction and effacement together with the mouse podocyte cell line shrinkage and the loss of cell contact.The above time point α-actin-4 mRNA expressions between the 2 groups were compared,and there was no significant difference in 8 h (P ＞ 0.05),but significant difference was found in 24 h,48 h,α-actin-4 higher mRNA expression,with statistical significance(all P ＜0.01).α-actin-4 in the control group had thin filaments evenly distributed in the cytoplasm,but a radioactive distribution in foot process.In the experimental group,α-actin-4 pressure silk fiber was shorter,with disordered arrangement,and PAN stimulus after 24 h,α-actin-4 distribution in cytoplasm was decreased significantly,while cytoplasmic distribution was missing after 48 h.Conclusions The abnormal of distribution and mRNA expression of α-actin-4 is time-related to the PAN injury podocyte,and α-actin-4 is an important part of podocyte damage mechanism.

Objective To observe the effect of puromycin aminonucleoside(PAN) on the expressions of CD2associated protein (CD2AP) and phosphatidylinositol 3-kinase (PI3 K) p85 in induced podocytes,and to explore the significance of abnormal expression of CD2AP in apoptosis of podocytes in vitro.Methods Mouse podocytes were injected into the control group and the PAN treatment group (PAN group).The cells were cultured for 8 h,24 h and 48 h respectively.The podocyte morphology was observed by phase-contrast microscope.The apoptosis ratio of podocytes was determined with flow cytometry.The mRNA expression of CD2AP was detected by real time fluorescent quantitative polymerase chain reaction.The protein expressions of CD2AP and PI3K p85 were detected by Western blot,respectively.The distributions and the relationships between CD2AP and PI3K p85 in the podocytes were detected by confocal fluorescence microscopy.Results PAN treatment led to significant shrinkage of podocytes with decreasing distribution tendency,podocyte foot process retraction as well as effacement and loss of cell contact.Compared with the control group,the apoptosis rate was increased at 24 h and 48 h[(7.52 ± 1.35)％,(17.09 ±2.53)％ vs (4.32 ±0.81％,(6.81 ± 1.34)％] in PAN group and the differences were significant (t =4.98,8.79,all P ＜ 0.05) ;CD2AP mRNA expression tended to decrease at each time point,and the differences were significant (0.34 ± 0.12,0.46 ±0.21,0.47 ± 0.10 vs 0.62 ± 0.23,0.97 ± 0.14,0.96 ± 0.23),and there were statistical differences (t =2.64,4.95,4.79,all P ＜ 0.05) ;CD2AP and PI3K p85 protein expressions tended to decrease at 24 h (0.36 ± 0.12,0.61 ± 0.20 vs 0.64 ±0.23,0.97 ±0.31) and 48 h(0.27 ± 0.15,0.48 ± 0.20 vs 0.51 ± 0.20,0.84 ± 0.35),and the differences were significant(t =2.64,2.31,2.35,2.40,all P ＜ 0.05).In the control group,the distribution of CD2AP was uniformly filamentary structure in cytoplasm and nucleus of podocytes,after PAN treatment the distribution of CD2AP changed to discontinuous coarse granular concentrated in the perinuclear.Immunocyte fluorescence showed that the distribution and the relationships between CD2AP and PI3K p85 in the podocytes became abnormal.Conclusions When the apoptosis of podocytes was induced by PAN,the expression and distribution of CD2AP were abnormal.CD2AP may play an important role in the survival of podocytes though the PI3K signaling pathway.

ObjectiveTo investigate the value of combined digital mammography and Color Doppler Mammasonography in the diagnosis of the breast carcinoma with ROC analysis.MethodsFifty female patients with breast carcinoma and fifty female patients with breast benign lesions confirmed by means of surgery and histopathological examinations were studied.All patients were examined by Digital Mammagraphy and Color Doppler Mammasonography.Images of all patients were sorted and divided into three groups,Digital radiographs of all patients was the first group,and the second group was Color Doppler photographs,and the third was included the above two photographs.Each group was evaluated respectively by three radiologists independently.The diagnostic result of breast carcinoma or benign lesions were investigated with a five point confidence-rating scale:(1)definitely positive;(2)probably positive;(3)indeterminate;(4)probably negative,and(5)definitely negative.ROC analysis was made with SPSS 11.0 software.The differences of the area under the curve for each group was made by Z test.ResultsThe average number of breast carcinoma diagnosed by three radiologists with digital mammography,Color Doppler Mammasonogaraphy and both of them was 42,38,and 48,respectively;and the average number of breast benign lesions was 40,43,and 47,respectively.The sensitivity and specificity for digital mammography was 84%(42/50),80%(40/50),and they were 76%(38/50),86%(43/50)for Color Doppler;and 96%(48/50),94%(47/50)for two modes combination.ROC analysis indicated that the average area under the curve was 0.852 for Digital Mammagraphy,0.825 for Color Doppler,and 0.952 for both combined.For digital mammography combined with Color Doppler Mammasono-garaphy on diagnosis breast carcinoma,the area under the curve(Az)was significally larger than that for digital mammography(Z1=2.183,P＜0.01),and also larger than that for Color Doppler Mammasono-garaphy(Z2=2.581,P＜0.01), respectively.Conclusions Among three medical imaging modes on diagnosing breast carcinoma,the diagnostic value of digital mammography combined with Color Doppler Mammasono-garaphy was the highest. It was helpful on detecting and diagnosing breast carcinoma early.

Objective To observe the expression of mRNA of podocin,nephrin,CD2AP and α - actin - 4 in Doxorubicin - induced nephrotic(ADN)rats,and explore the possible mechanisms of podocyte molecule during the de-velopment of proteinuria. Methods Forty - eight Sprague - Dawley(SD)rats were divided into ADN model group(in-jected with 6. 5 mg/ kg Doxorubicin in tail vein,n = 24)and control group(injected with saline solution in tail vein,n =24). After the nephropathy model was established,6 rats were killed at the end of 1st ,2nd ,4th ,6th week in each group. The changes of the following indicators were observed:(1)24 - hour urinary protein,serum albumin and cholesterol were detected;(2)mRNA expression of nephrin,podocin,CD2AP and α - actin - 4 in cortex of kidney were examined by real time fluorescence quantification PCR. Results The model group came out massive proteinuria(15. 66 ± 1. 50) mg/ 24 h,(45. 98 ± 1. 45)mg/ 24 h,(65. 58 ± 4. 68)mg/ 24 h,(82. 83 ± 8. 43)mg/ 24 h in 1,2,4,6 weeks respec-tively,hypoalbuminemia(27. 4 ±2. 5)g/ L,(23. 6 ±2. 9)g/ L,(20. 6 ±1. 5)g/ L,(6. 9 ± 2. 3)g/ L in 1,2,4,6 weeks respectively and hypercholesterolaemia(2. 00 ± 0. 25)mmol/ L,(2. 16 ± 0. 44)mmol/ L,(4. 02 ± 0. 81)mmol/ L, (7. 54 ± 1. 12)mmol/ L in 1,2,4,6 weeks respectively,and the differences of proteinuria,plasma albumin and total cholesterol compared with control group at each time point had statistical significance(all P ﹤ 0. 01). Compared with the control group,podocin mRNA expression in the model group decreased at the end of 1st week(10. 56 ± 3. 62),de-creased significantly at the end of 2nd week(20. 44 ± 9. 03),and decreased at the end of 4th week(2. 19 ± 0. 18)com-pared with the control group;nephrin mRNA expression decreased at the end of 1st week(2. 41 ± 1. 10)and reached to the peak value,decreased at the end of 4th week(0. 52 ± 0. 18);CD2AP mRNA expression did not change significantly in the 1st week(4. 17 ± 0. 79),increased at the end of 2nd week(6. 74 ± 1. 53),reached to the peak value at the end of 4th week(6. 91 ± 1. 13),but did not change significantly at the end of 6th week(4. 04 ± 0. 82);α - actin - 4 mRNA ex-pression did not change significantly at the end of 1st week(1. 75 ± 0. 48),decreased at the end of 2nd week(2. 01 ± 0. 55),reached to the peak value at the end of 4th week(2. 24 ± 0. 81),but did not change significantly at the end of 6th week(1. 39 ± 0. 18). Compared with the control group,the difference had statistical significance( all P ﹤ 0. 05). Conclusion The abnormal expression of podocyte molecules mRNA in ADN rats may be an important molecular mechanism in the development of proteinuria.

High expression of Fightless I (FLII) is associated to multiple tumors. Based on our previous study that FLII might be involved in the nuclear export, we assessed the possible interaction of FLII with the nuclear envelop associating proteins Importin β and Nup88. We first constructed GST-FLII, GST-LRR recombinant plasmids and transformed them into the Rosetta strain to produce GST-FLII, GST-LRR fusion protein. After purification of these proteins, GST-pull down, as well as co-immunoprecipitation, were used to test the interaction of FLII with Importin β and Nup88. FLII interacted with Importin β and Nup88, and FLII LRR domain is responsible for these interactions. Thus, FLII may play a role in nuclear export through interaction with Importin β and Nup88.
Humans ; Microfilament Proteins ; metabolism ; Nuclear Pore Complex Proteins ; metabolism ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Recombinant Fusion Proteins ; metabolism ; beta Karyopherins ; metabolism

Agrimonia pilosa is one of the traditional Chinese herbal medicines for 2000 years history. Due to its various pharmacological effects, it is widely used in the clinical treatment of tumors and other diseases. Presently malignancy has become an important factor threatening human health that the anti-tumor effect of Agrimonia pilosa has attracted great attention. The author reviewed its anti-tumor effect mechanism by articles arrangement of experimental study, clinical study and experience introduction. A series of study revealed 3 mechanisms: blocking cell cycle, inducing apoptosis and strengthening the cellular immunity. As a kind of traditional Chinese herbal medicine which has anti-tumor effect, the Agrimonia pilosa should be reasonably applied according to the related research and previous experience.

Objective To analyze the 14 indicators of on-site evaluation at 18 hospitals under Beijing hospital authority in 2016 , and to provide technical support and reference for further optimization of medical quality and service .Methods According to the "Beijing Municipal Administration of Hospital 2016 On-site Evaluation Indicators", the results were analyzed using the fuzzy combined method of TOPSIS and rank sum ratio , and the ranking of the indicators was sorted .Results TOPSIS method and rank sum ratio weighted fuzzy joint analysis showed that the top three indicators from high to low were medical technology management , patient identification and verification system , clinical care service management;and the last three indicators were hospital-acquired infection monitoring , medical malpractice reporting and management , critical value report and disposal .Conclusions Hospitals are recommended to strengthen their exchange , popularize the concept of continuous improvement , the use of management tools to solve practical problems , and further improve the hospital medical quality and service quality .

Objective:To conduct data mining on hospital accreditation results for inspirations and clues in hospital management.Methods:The Apriori function contained in the arules package of R software was used to extract the association rules. This practice aimed to analyze association rules of the accreditation results of 56 tertiary hospitals which were made based on tertiary hospital accreditation standard(2011)and Detailed Rules from 2017 to 2019; to explore the correlation between the clauses, and analyze the inspirations for hospital management.Results:A total of 6 566 138 and 247 rules were generated for all clauses and core clauses, receptively. The top 10 rules sorted by lift were selected as strong association rules. Among them, the minimum lift of all clauses and core clauses was 1.41 and 1.53, respectively. There was a strong correlation between the establishment of a quality and safety management team in the hospital and the development & implementation of relevant regulations by the hospital. There were strong correlations among emergency service procedure and regulations, patient rights to know, responsibility system by the first one to receive a complaint, medical safety(adverse)event reporting, development & implementation of antimicrobial management system, as well as training for prevention of multi-drug resistance infection measures, and multi-drug resistance infection hospital infection control system.Conclusions:This study suggested that hospital management should highlight the correlation between regulations development and staffing, that between indicator systems and monitoring feedback systems, and that between indicators in different sectors in the medical process. These correlations can be used as management clues and inspirations for hospital management.

Objective To prepare 131I-anti-neuropilin-2-monoclonal antibody (131I-anti-NRP-2-mAb),and investigate its biodistribution and imaging in nude mice bearing xenografted lung adenocarcinoma,in order to evaluate its feasibility as an imaging agent targeting to NRP-2 positive tumors.Methods (1)131I-anti-NRP-2-mAb was prepared by Chloramine-T method under the optimum labeling conditions,then the labeling efficiency,radiochemical purity and stability were determined in vitro.(2) The binding fraction and receptor binding affinity of 131I-anti-NRP-2-mAb were measured in A549 human lung cancer cells by cell uptaking and binding experiments.(3) The A549 tumor-bearing mice were randomly divided into 4 groups with direct sampling method and were sacrificed at 6,24,48,and 72 h,respectively,after tail intravenous injection of 0.37 MBq 131I-anti-NRP-2-mAb.The distribution was measured,and the ratios of tumor/muscle (T/M) and tumor/blood (T/B) were calculated.(4) Gamma imaging was performed in 6 mice,including 3 in the competitive inhibition control group (injected with 3.7 MBq 131I-anti-NRP-2-mAb and 100 μg atniNRP-2-mAb),at 6,24,48,and 72 h post-injection to observe the radioactivity in tumor.Two-sample t test was used for data analysis.Results (1) The labeling yield and radiochemical purity of 131I-anti-NRP-2-mAb were (94.69 ± 3.63) ％ and (98.56± 0.48) ％,respectively.The radiochemical purity was more than 85％ after incubating in phosphate-buffered solution at room temperature for 72 h.(2) At 60,120,180 and 240 min post-injection,the binding ratios of 131I-anti-NRP-2-mAb in A549 cells were (3.95±0.18)％,(5.19±0.65) ％,(6.60± 0.36) ％ and (5.58± 0.63) ％,respectively.When excessive anti-NRP-2-mAb were added,the binding ratios were reduced to (0.94±0.31)％,(1.12±0.17)％,(1.24±0.25)％ and (1.04±0.18) ％,respectively,which were significantly lower than those of non-inhibited group (t values:9.89-19.66,all P＜0.05).131I-anti-NRP-2-mAb bound to NRP-2 with high affinity half maximal inhibitory concentration (IC50 =(410.8±1.2) nmol/L).(3) Biodistribution study demonstrated that the T/M and T/B ratios increased with the time extension and were 3.83±0.18 and 1.10±0.20,respectively,at 72 h post-injection.(4) Gamma imaging studies revealed that 131I-anti-NRP-2-mAb could clearly identify A549 tumors 6 h post-injection,especially at 48 h post-injection.Tumors were not observed clearly in competitive inhibition control group.Conclusion 131I-anti-NRP-2-mAb has been successfully prepared,and it could target to NRP-2 specifically.