Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.

ObjectiveTo investigate the role of reducing the door-to-needle time for patients with acute ischemic stroke based on the quality improvement program of PDCA cycle.MethodsConsecutive patients with acute ischemic stroke admitted to hospital were registered prospectively from January 1, 2016 to September 30, 2016.Questionnaires and time tracking method were used to investigate the door-to-needle (DNT) and its influencing factors.PDCA cycle method was used to improve the stroke channel workflow and the changing trend of DNT was analyzed.ResultsA total of 71 patients with acute ischemic stroke were enrolled.After 3 PDCA cycles, DNT (median, interquartile range) from 100.0 min (65.5-127.0 min) reduced to58.0 min (45.5-80.0 min) (Z=11.689, P<0.001), the proportion of the patients with DNT ≤60 min increased from 19.05% to 60.00% (χ2=7.893, P=0.019).Conclusions The quality improvement program of PDCA cycle may effectively reduce the time of DNT in patients with acute ischemic stroke.

Background Induced pluripotent stem cells (iPSCs)can differentiate into various types of somatic cells without causing ethical controversy and immune rejection in clinical activity,which is similar to differentiation ability of embryonic stem cells.So,iPSCs may be used as seed cells for tissue engineering corneal endothelial reconstruction.Objective The present study was to survey the morphologic change of iPSCs after coculture with corneal endothelium cells(CECs) under the atomic force microscopy(AFM).Methods Rabbit CECs and human MMC-iPSCs were isolated and cultured respectively.The iPSCs were identified with the marker by immunochemistry.iPSCs passaged for 7 days were then cultured with 60％ confluent CECs to establish the co-culture model.The surface morphology and cellular membrane ultrastructure of differentiated iPSCs after induced by CECs were examined by AFM combination with inverted microscope,and compared with CECs and undifferentiated iPSCs.Results Thelengthand width were(66.93±10.48)μm and (44.85 ± 8.14) μm in CECs,(12.51±1.40)μm and (10.93 ±1.69) μm in uninduced iPSCs,and(36.12±10.29) μm and(31.53±9.65)μm in CECs-induced iPSCs.Both the length and width values of CECs-induced iPSCs were statistically bigger than those uninduced iPSCs,with significant differences between them (P＜0.05),but no significant difference was seen in the width valne of CECs-induced iPSCs in comparison with CECs(P＞0.05).The convex structure of CECs cytomembrane surface showed the digitation in shape with the size and height(2.11 ± 1.03) μm and (115.68±92.08) nm respectively,and the concave structure of cytomembrane surface of CECs was fenestrae-like depression and the size was (1.49 ± 0.65) μm.The numerical valuc of mean square root roughness (Rq)and average roughness (Ra)of cytomembrane surface of CECs were(39.20±7.82)nm and (30.37±5.32)nm respectively.The convex surface of cytomembrane of iPSCs was granular-like in shape with size and height(0.39±0.22)μm and(13.11±9.18)nm respectively.The concave surface of cytomembrane of iPSCs was worm-eaten-like concave with the size(0.34±0.18)μm.The numerical value of Rq and Ra of geometrical parameters of cytomembrane surface of iPSCs were (26.60 ± 4.93)nm and (9.97 ± 3.78) nm respectively.The convex surface of cytomembrane of induced iPSCs was digital-like in shape with the size and height (1.91±0.76) μm and(106.55±77.27) nm respectively.The concave surface of cytomembrane of induced iPSCs was fenestrae-like depression and the size of concave was(1.6l±1.25) μm.The numerical value of Rq and Ra on surface of cytomembrane of induced iPSCs was (57.33± 12.80) nm and (43.63± 11.17) nm respectively.The numerical values of the size and height of convex,the size of concave,Rq and Ra on surface of cytomembrane in induced iPSCs were statistically bigger than in iPSCs(P＜0.05)and were not significant differences in comparison with CECs (P＞0.05).Conclusions Morphology of iPSCs translate toward the CECs after induce for 7 days under the AFM.This outcome lays the foundation for further study on iPSCs.

Objective To investigate the influence of anti-depression therapy on cardiac function and short-term prognosis in elderly patients with congestive heart failure (CHF) and the safety and effectiveness of fluoxetine in these patients.Methods Ninety six elderly hospitalized patients with CHF accompanied with depressive disorder were randomly divided into two groups:fluoxetine (20mg once daily)and placebo treated groups based on the routine cardiac drug therapy for 3 months. Results The prevalence rate of depressive disorder in elderly patients with CHF was 31.1%.The length of stay of fluoxetine group was shorter than that of placebo group.The post-treatment depressive exponents of self-rating depression scale(SDS)in fluoxetine group (51.39%?8.63)was lower than those in pretreatment and in placebo group.The improvement of cardiac function in fluoxetine group(53.4%?4.5%) was much better than that of placebo group (P

Animals ; Drugs, Chinese Herbal ; pharmacology ; Liver Cirrhosis, Experimental ; enzymology ; Male ; Pancreatic Elastase ; biosynthesis ; genetics ; Rats ; Rats, Wistar

INTRODUCTION: A second-tier rapid response team (RRT) is activated for patients who do not respond to first-tier measures. The premise of a tiered response is that first-tier responses by a ward team may identify and correct early states of deterioration or establish goals of care, thereby reducing unnecessary escalation of care to the RRT. Currently, utilisation and outcomes of tiered RRTs remain poorly described. METHODS: A prospective observational study of adult patients (age ≥18 years) who required RRT activations was conducted from February 2018 to December 2019. RESULTS: There were 951 consecutive RRT activations from 869 patients and 76.0% patients had a National Early Warning Score (NEWS) ≥5 at the time of RRT activation. The majority (79.8%) of patients required RRT interventions that included endotracheal intubation (12.7%), point-of-care ultrasound (17.0%), discussing goals of care (14.7%) and intensive care unit (ICU) admission (24.2%). Approximately 1 in 3 (36.6%) patients died during hospitalisation or within 30 days of RRT activation. In multivariate analysis, age ≥65 years, NEWS ≥7, ICU admission, longer hospitalisation days at RRT activation, Eastern Cooperative Oncology Group performance scores ≥3 (OR [odds ratio] 2.24, 95% CI [confidence interval] 1.45-3.46), metastatic cancer (OR 2.64, 95% CI 1.71-4.08) and haematological cancer (OR 2.78, 95% CI 1.84-4.19) were independently associated with mortality. CONCLUSION Critical care interventions and escalation of care are common with second-tier RRTs. This supports the need for dedicated teams with specialised critical care services. Poor functional status, metastatic and haematological cancer are significantly associated with mortality, independent of age, NEWS and ICU admission. These factors should be considered during triage and goals of care discussion.
Adolescent ; Adult ; Aged ; Critical Care ; Hospital Mortality ; Hospital Rapid Response Team ; Humans ; Prospective Studies ; Tertiary Care Centers

OBJECTIVETo study the mechanism of Fuzheng Huayu (FZHY) recipe against pulmonary fibrosis relating to MMP-2 activity and type IV collagen expression at lung tissue.

METHODThe pulmonary fibrosis model was induced by intratracheal instillation with bleomycin once in rats. The models were divided into 3 groups: model control, FZHY recipe treated, and methylprednisolone (Solu-Medrol) treated group, each group was of 14 model rats. Normal control group with 10 rats was intoxicated with the same amount of saline. From the second day of intoxication, FZHY recipe treated group orally took FZHY recipe at the dosage of 4.6 g x kg(-1) rat wt, methylprednisolone treated group received intraperitoneal injection with 15 mg x kg(-1) rat wt of methylprednisolone, while model and normal controls took the same volume of saline, 1 time each day and lasting for 4 weeks. Lung and body weights were weighed and the lung/body ratio was calculated. Collagens deposition was check with Masson stain, and lung hydroxyproline (Hyp) content was assayed with Jamall's method. Protein expressions of MMP-2/9 and type IV collagen at lung tissue were analyzed with Western blot and of MMP-2/9 activities by gelatin zymography.

RESULTCompared to normal rats, the model control rats had a high lung/body ratio, remarkable collagen deposition, increased Hyp content and the expressions of type IV collagen, MMP-2 and MMP-9 protein at lung tissue, increased MMP-2 activity, in particular active MMP-2 activity, but decreased MMP-9 activity. Compared to model control, FZHY recipe and methylprednisolone obviously attenuated pulmonary collagen deposition, decreased lung/body ratio and Hyp content, downregulated MMP-2 protein expression and activity, in particular active MMP-2 activity, and FZHU recipe had some better actions than methylprednisolone on lung/body ratio, type IV collagen expression and active MMP-2 activity. But both drug groups had no influence on MMP-9 protein expression and activity.

CONCLUSIONFZHY recipe has a good action against experimental pulmonary fibrosis and its mechanisms are associated with the inhibition of MMP-2 protein and activity, and with the inhibition of over expression of type IV collagen at lung tissue.

Animals ; Bleomycin ; Collagen Type IV ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hydroxyproline ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Plants, Medicinal ; chemistry ; Pulmonary Fibrosis ; chemically induced ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Aged ; Beijing ; China ; Emergency Service, Hospital ; Hospitalization ; Humans ; Temperature

OBJECTIVETo explore the mechanism of hypoalbuminemia in patients with severe sepsis.

METHODSI(125)-labeled albumin was administered intravenously to 10 health volunteers and 10 patients with severe sepsis. Blood samples were taken at 0, 1, 2, 4, 8, 12, 24 hours and 2, 3, 4, 5, 6, 7, 9, 11, 13, 15, 18, 22, 25 days for the measurement of the dose of gamma-radiation and the curve of concentration and time. Then the half-life time (t(1/2)), apparent volume of distribution (V(d)) and transportation rate (K(12)) from center compartment to side compartment of albumin were calculated.

RESULTSThe half-life time in septic group was obviously shorter than that in control group (8.2 +/- 1.4 vs. 12.5 +/- 1.7, P < 0.01). The transportation rate in the septic group was higher than that in the control group [(4.4 +/- 1.9) x 10(-2)/h vs. (2.4 +/- 0.6) x 10(-2)/h, P < 0.05]. There was no significant difference in apparent volume of distribution between the two groups.

CONCLUSIONSIn patients with severe sepsis, the distribution rate of albumin from vessel to tissue was obviously increased and the decomposition rate of albumin was markedly improved.

Adult ; Aged ; Female ; Half-Life ; Humans ; Kinetics ; Male ; Middle Aged ; Sepsis ; metabolism ; Serum Albumin ; metabolism

OBJECTIVETo screen and identify the possible existence of natural antisense transcript (NAT) within the mouse neocortex.

METHODSSixty-three cerebral cortex layer-specific genes were screened by bioinformatics prediction in mice, among which 31 mice with potential NATs were screened. NAT was identified using reverse transcription polymerase chain reaction (RT-PCR) and then cloned in pGEM-T Vector System for sequencing.

RESULTSAmong 31 genes predicted using bioinformatics, 8 were proved to be NAT positive by RT-PCR.

CONCLUSIONSNATs exist in the mouse neocortex tissue during the development of cerebral cortex. NATs may influence mouse cortical development by regulating the related coding genes.

Animals ; Cell Line ; Cerebral Cortex ; Mice ; Molecular Sequence Data ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics