Objectives: To improve the procedures for establishment of focal cerebral ischemic rat models with the intraluminal suture method and to elevate the survival rate of the models. Methods: Sixty male SD rats (weight, 250-270 g) were randomly allocated into 3 groups (n =20 in each group). Control group used the conventional surgical method, the pterygopalatine arteries were exposed and ligated during the procedure, and the rats had free access to food after the procedure; gavage group used the same method as the control group, but postoperative gavage was performed; protective group did not ligate the pterygopalatine arteries, but the carotid arteries and vagus nerves were carefully separated. Trying to avoid stretching the vagus nerves during the procedure and postoperative gavage was performed. The body weight of the rats in each group was weighed continuously before and after modeling, and the neurological deficit scores were measured; the survival rate of rats in each group was calculated at day 3, 14, and 28, respectively after modeling; the brain infarct volume in each group was calculated at day 28 after modeling. Results: Circled digit oneThe mean time of model production was 19.5 min in the protective group, and it was shorter than that in the control and gavage groups. The differences were statistically significant (P < 0.05); Circled digit twoAt day 3 after modeling, the survival rate of the rats in the protective group was higher than that in the control and gavage groups. The differences were statistically significant (P < 0.05); Circled digit threeAfter modeling, the body weight of the rats in the 3 groups were all decreased. The body weight of the rats in each group began to increase from day 7 after modeling. The body weight of the rats in the protective group was all higher than that in the control and gavage groups at day 7, 14, 21, and 28 after the procedure. The differences were statistically significant (P < 0.05); Circled digit fourthere were no significant differences comparing the neurological deficit scores and cerebral infarction volume among the 3 groups. Conclusion: The improved procedures could shorten the time of model production, regain body weight quickly, improve the survival rate effectively after establishment of the cerebral ischemic rat models.

Cardiovascular Diseases ; Humans ; MicroRNAs

Adrenal Cortex Hormones ; therapeutic use ; Adrenergic beta-Agonists ; therapeutic use ; Anti-Asthmatic Agents ; therapeutic use ; Asthma ; drug therapy ; Humans ; Medication Adherence

Objective To explore the influence of carvedilol combined with diuretics on BNP and cardiac function of patients with mild to moderate cardiac insufficiency.Methods87 cases of patients with mild to moderate cardiac insufficiency treated in Zhejiang Greentown Cardiovascular Hospital from June 2014 to June 2016 were divided into the groups randomly.The control group of 44 cases were given carvedilol treatment, and the study group were combined with spironolactone treatment on the basis of the control group, with the total course for a month.Clinical effects were contrasted after a month of treatment in the two groups, in order to explore the influence of carvedilol combined with diuretics on BNP and cardiac function of patients with mild to moderate cardiac insufficiency.ResultsBefore treatment, there was no significant difference in serum BNP level between the control group and the study group.After treatment, serum BNP level significantly decreased when compared with those before treatment in the two groups (P< 0.05), which of the study group was significantly lower than that in the control group(P< 0.05).Before treatment, there were no significant differences in the levels of cardiac function indexes, and after treatment, heart rate, EF (%), FS (%), LVEDD (mm) the four indicators significantly improved when compared with those before treatment, and heart rate, LVEDD (mm) levels were significantly lower than those in the control group(P<0.05), and EF (%), FS (%) levels were significantly higher thatn those in the control group(P<0.05).After treatment, the significant effective rate (73.91%) and total effective rate (95.35%) in the study group were significantly higher than that of (56.82%) and(86.36%) in the control group (P<0.05).After treatment, body function, life satisfaction, mental health, social adjustment the four parts of quality of life index scores were significantly higher than those in the control group(P<0.05).ConclusionIt has obvious improvement effect of carvedilol combined with diuretics on BNP and cardiac function of patients with mild to moderate cardiac insufficiency.It is worthy of clinical application.

Objective To discussion the clinical study of milrinone injectionon on acute heart failure with poor effect of digitalis and diuretics.Metheds 90 cases of acute heart failure with poor effect of digitalis and diuretics patients from September 2014 to May 2016 in our hospital were selected,according to the random number table method is divided into control group and study group,45 cases in each groups,the control group was treated with conventional heart failure intervention program,the study group given milrinone injectionon on the basis of control group,two groups were treated for seven days.Serum neurohormonal factors,inflammatory factors and blood pressure were measured before and after treatment,and the clinical efficacy and complications were compared between two groups.Results Compared with before treatment,serum NT-proBNP,NE and ET-1 levels were decreased in two groups,NO content increased,the levels of serum TNF-α,IL-6 and Hs-CRP were decreased,the levels of LVEF and CI were increased,the levels of LVEDD and CTR were decreased,the differences were statistically significant(P<0.05).Compared with the control group,the levels of serum NT-proBNP,NE and ET-1 in the study group were lower,NO were higher,the levels of TNF-α,IL-6 and Hs-CRP in serum were lower,LVEF and CI were higher,LVEDD and CTR were lower,the differences were statistically significant(P<0.05).The effective rate of the study group was 86.67％,which was significantly higher than that of the control group(66.67％,P<0.05).All the patients were followed up and no cases were reported.There was no significant difference in the incidence of adverse reactions between the two groups.Conclusion Milrinone injectionon has a significant effect on acute heart failure with poor effect of digitalis and diuretics,can significantly reduce the neurotrophic factor and inflammatory factors,improve heart function,promote prognosis.

Background Aberrant proliferation of residual lens epithelial cells (LECs) is one of main causes of posterior capsular opacification (PCO).Researches indicated that Wnt3a signaling pathway promote proliferation of epithelial cells,but its effect on LECs is still unclear. Objective The present study was to investigate the effects of Wnt3a on proliferation of human LECs and its mechanism and to provide a new gene target in the prevention and treatment of PCO. Methods Human LECs line (SRA01/04 cells ) was cultured and then incubated to 6-well plate at the density of 4×105/well.A human Wnt3a cDNA expressing vector targeted human LECs was constructed to increase the Wnt3a expression in SRA01/04 cells,and pcDNA3-HA expression vector was used as the control group.The expression of Wnt3a was identified by Western blot assay after transfected.The growth and proliferation of SRA01/04 cells were detected by MTT and flow cytometry (FCM).The expressions of β-catenin,cyclin D1 and c-myc in the cells were detected by Western blot assay.β-Catenin expression was localized using immunofluorescence assay,and the expression and localization of proliferating cell nuclear antigen ( PCNA ) were analyzed by immunocytochemistry for the exploration of the active mechanism of Wnt3a to proliferation of LECs.Results Human Wnt3a cDNA expression vector was designed successfully and transiently transfected to SRA01/04 cells,and Wnt3a/SRA01/04 cells and pcDNA3-HA/SRA01/04 cells were obtained.The expression of Wnt3a was verified in the Wnt3a transfected group compared with the control group.MTT indicated that the cell proliferating rate was significantly different between the Wnt3a transfected group and the control group ( Fgroup =15.235,P =0.005 ;Ftime =369.677,P =0.000),and that in various time points after transfected was significantly different (t =20.843,P=0.001 ;t =26.214,P＜0.001 ;t=25.177,P=0.001 ;t =35.516,P＜0.001 ;t =615.056,P＜0.001 ).The proportion of SRA01/04 cells in G1 phase was 51.74％ in the Wnt3a cDNA transfected group,with a significantly decrease in comparison with 79.44％ of the control group.However,the proportion of SRA01/04 cells in S phase in the Wnt3a cDNA transfected group was higher than that of the control group (36.23％ versus 12.34％ ).The positive expression rate of PCNA protein in SRA01/04 was (47.00％ ±7.58％ ) in the Wnt3a cDNA transfected group and ( 16.00％ ±3.61％ ) in the control group with a significant difference between them (t =8.256,P＜0.01 ).After 48 hours of transfection of the Wnt3a cDNA,the expression amount of β-catenin proteins was higher and the immunofluorescence was stronger in cell nucleus,and the expressions of cyclin D1 and c-myc proteins were elevated in Wnt3a/SRA01/04 cells. Conclusions The overexpression of Wnt3a activates the Wnt/β-catenin signaling pathway and downregulates the expression of a subset of target genes,including cyclin D1 and c-myc,which plays an important role in promoting the proliferation of human LECs.

Objective: To study the chemical constituents from Corydalis Rhizoma in Pan'an county as characteristically regional traditional Chinese medicinal materials. Methods: The constituents were separated and purified by chromatography on silica gel and Sephadex LH-20 columns, as well as the chemical structures of alkaloids were determined by physicochemical properties and spectral data analyses. Results: Twenty alkaloids were obtained and identified as tetrahydrocoptisine (1), tetrahydrocolumbamine (2), tetrahydropalmatine (3), corybulbine (4), isocorybulbine (5), 8-trichloromethyl-7, 8-dihydrocoptisine (6), corydaline (7), 8-oxocoptisine (8), (-)-corypalmine (9), dehydrocorydaline (10), 13-methylpalmatrubine (11), oxoglaucine (12), protopine (13), noroxyhydrastinine (14), tetrahydroberberine (15), didehydroglaucine (16), pontevedrine (17), coptisine (18), palmatine (19), and berberine (20). Conclusion: Compounds 6, 9, and 16 are isolated from Corydalis Rhizoma for the first time. All 13C-NMR and 1H-NMR data of compounds 6 and 16 are assigned for the first time using 2D NMR.

Humans ; Eosinophilia/diagnosis* ; Thrombosis/etiology* ; Heart Diseases/complications*

This article introduces the general concepts and methods of sample size estimation and testing power analysis. It focuses on parametric methods of sample size estimation, including sample size estimation of estimating the population mean and the population probability. It also provides estimation formulas and introduces how to realize sample size estimation manually and by SAS software.

Objective:To study the isolation, culture and identification of the TMJ cells and to observe the biological characteristics of cultured fibrochondrocytes. Methods:The TMJ discs were dissected from two 1 month goats under sterile conditions and were digested with collagenase. The cells were collected. Morphological changes and attachment efficiency were constantly observed under phase-contrast microscope. Immunohistochemical staining for type I collagen as well as toluidine blue staining were performed. Ultrastructures of the TMJ cells were observed under transmission electron microscope. Results: Most of the primary fibrochondrocytes presented a short spindle-shape while the rest showed polygon-shape. On the 7th day, the perliferating fibrochondrocytes started to contact each other to form a monolayer covering the bottom of the incubation disc. Immunohistochemical staining of type I and toluidine blue staining exhibited positive results. The fibrochondrocytes cytoplasms were rich in mictochondria and endoplasm reticulum. Conclusion: The fibrochondrcytes isolated from one-month-old goat TMJ disc have good proliferation ability in vitro and cells from passage 1 to 3 might be used as seed cells for TMJ disc tissue engineering.