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 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.

Endoscopic ultrasonography (EUS)is routinely performed before endoscopic submucosal dissection (ESD)for treatment of upper gastrointestinal stromal tumors.However,when a miniprobe sonography (12,15 and 20 MHz)was used,the size of tumor revealed by EUS was often inconsistent with what it actually was,which might increase the difficulty of ESD and the risk of perforation and massive bleeding.Aims:To investigate the value of standard probe (5 and 7.5 MHz)EUS in detecting the size of upper gastrointestinal stromal tumors before ESD.Methods:Clinical data of patients who were suspicious of esophageal and gastric stromal tumors by gastroscopy and EUS from Jan.2012 to Oct.2014 at the Renmin Hospital of Wuhan University were collected.Of them,195 cases treated by ESD were retrospectively analyzed.Results:Of 195 cases treated by ESD,37 cases diagnosed by standard probe EUS and 108 cases diagnosed by miniprobe EUS were confirmed as stromal tumors by pathology.Fourteen cases were failure for ESD and then transferred to surgical treatment,one was due to misjudgement of the origin of tumor by standard probe EUS and 9 were due to misjudgement of the size of tumor by miniprobe EUS.The misjudgement rate of standard probe EUS was lower than that of miniprobe EUS with an insignificant difference (2.7%vs.8.3%,P>0.05).In 9 cases misjudged by miniprobe EUS, the size of tumor presented by miniprobe EUS was significantly smaller than its real size [(1.22 ±0.51)cm vs.(3.97 ±1.06)cm,P<0.01].ESD was avoided or terminated in 3 cases because of the accurate estimation of tumor origin, structure and blood flow by standard probe EUS.Conclusions:For patients who are going to receive ESD for suspected upper gastrointestinal stromal tumors,it would be best to select standard probe EUS to detect the size,origin and blood flow of the tumor before ESD.It will decrease the risk and improve the success rate of ESD.

Aim To investigate the effects of ursolic acid from loquat leaves on proliferation inhibition and expression of PPAR-γ 、TGF-β1 in rat hepatic stellate cells, so as to explore the mechanism of anti-hepatic fibrosis of UA.Methods HSC-T6 cells were randomly divided into blank group, rosiglitazone control group, the low, medium and high concentration of UA group to detect the cell proliferation inhibition by CCK-8 after 24,48,72 h.The content of type collagen Ⅰ in cell culture supernatant of each group was examined by ELISA.The expression of PPAR-γ mRNA, TGF-β1 mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR.Effects on HSC-T6 PPAR-γ and TGF-β1 protein from each group were detected by immunocytochemical method.Results CCK-8 results showed that the inhibitory rate of UA on cell proliferation increased with the prolongation of drug action time(P<0.01).ELISA results showed that with the increase of the concentration of UA, the content of type Ⅰ collagen content decreased(P<0.01).Real-time PCR results showed that with the increase of the concentration of UA, and the expression of PPAR-γ mRNA increased, the expression of TGF-β1 mRNA decreased(P<0.01).The results of immunocytochemistry showed that the expression of PPAR-γ protein was increased, and the expression of TGF-β1 protein was decreased with the increase of the concentration of UA(P<0.01).All effects mentioned above were dose-dependent.Moreover, the effects in the high concentration groups were stronger than those in control group.Conclusion UA can inhibit the proliferation of HSC-T6 cells, Which may be associated with the up-regulation of PPAR-γ expression and the down-regulation of TGF-β1 expression.

ObjectiveTo estimate the applications of ADC value and rADC value in the diagnosis of nodular lesions of breasts.Methods Fifty-two cases with 66 nodular lesions of breasts confirmed by histopathology underwent diffusion-weighted magnetic resonance imaging.Three b values (0,800 and 1000 s/mm2) were applied.The mean ADC values of the breast nodules,the ADC values of ipsilateral breast( rADC1 )and ADC values of contralateral breast (rADC2 )were respectively measured.The independent-samples t-test and chi-square test were used for statistical analyses.ResultsOf the 52 patients,there were 18 patients with infiltrating ductal carcinoma and 34 patients with fibroadenoma.50 patients with 64 lesions were examined by DWI.( 1 ) at b = 800 s/mm2,the mean ADC values of malignant nodules [ ( 1.01 ±0.09) × 10-3 mm2/s],rADC800-1 (0.52 ±0.07)and rADC800-2 (0.51 ±0.06) were lower than that of the benign nodules [ ADC value = ( 1.54 ± 0.28 ) × 10 -3 mm2/s,t = 8.217,P ＜ 0.01 ; rADC800-1 =0.77 ±0.15,t =9.339,P＜0.01 ; rADC800-2 =0.76 ±0.14,t = 10.394,P ＜0.01 ].The one-side upper limits of 95％ medical reference value of mean values of infiltrating ductal carcinoma were adopted as the threshold point to distinguish the malignant from the benign.The threshold value of breast malignant nodule ADC,the rADC800-1 and rADC800-2 were respectively 1.05 × 10-3 mm2/s,0.55 and 0.53.The sensitivities of the three methods were 75.0％,65.0％ and 60.0％ ; the specificities were 100.0％,95.7％ and 97.8％ ;the positive predictive values were respectively 100.0％,86.7％ and 92.3％ ; the negative predictive values were 90.2％,86.3％ and 84.9％; the diagnosis accordance rates were respectively 92.4％,86.4％ and 86.4％.( 2 ) at b = 1000 s/mm2,the mean ADC values of malignant nodules [ ( 0.93 ± 0.08 ) ×10-3 mm2/s],rADC1000-1 (0.53 ±0.09) and rADC1000-2 (0.52 ±0.07) were also lower than that of the benign nodules[ ADC value= (1.45 ±0.28) ×10-3 mm2/s,t=11.844,P＜0.01; rADC1000-1 =0.75 ±0.16,t=5.820,P ＜ 0.01 ; rADC1000-2 = 0.74 ± 0.15,t = 8.082,P ＜ 0.01 ].The threshold value points breast malignant nodule ADC,the rADC1000-1 and rADC1000-2 were respectively 0.97 × 10-3 mm2/s,0.58,0.55.The sensitivities were all 70.0％ ; the specificities were respectively 100.0％,95.7％ and 93.5％ ;the positive predictive values were 100.0％,87.5％ and 82.4％ ; the negative predictive values were 88.5％,88.0％ and 87.8％ ; the diagnosis accordance rates were 90.9％,87.9％ and 86.5％ respectively.There were no significant differences in specificities and the diagnosis accordance rates ( x2 = 1.232,2.263 ; P =0.942,0.812 ).Conclusions ADC value and rADC value are both important parameters of MRI in differentiating benign and malignant breast diseases.The study indicated that ADC value ( at b =800 s/mm2) was the most valuable parameter.

Background The multipotent differentiation features of induced pluripotent stem cells (iPSCs) offer a new option for cell replacement therapy of many clinical diseases.In ophthalmology,iPSCs are a good model in studying the pathogenic mechanism of degenerative ocular diseases.A better identification method for iPSCs is critical for analyzing the in vivo biological characteristics of iPSCs.Objective This study was to investigate the feasibility and stability of labeling iPSCs with quantum dots.Methods Human umbilical mesenchymal stromal cells-iPSC lines were cultured and amplified on matrigel,and the characteristics of iPSCs were evaluated by immunofluorescence.Different concentrations (5.0,7.5 and 10.0 nmol/L) of quantum dots with a CdSe/ZnS nuclear shell structure were used to label iPSCs after passaging and proliferation.The labeling outcome was observed with a three-dimensional deconvolution real-time live cells imaging system.The labeled iPSCs were subsequently cultivated,and then changes in fluorescence intensity were examined 7 days after the first and the second passaging of iPSCs.Results iPSCs were observed to grow in a clonal manner under the inverted microscope.The iPSC markers,OCT4 and Nanog,were detected by immunofluorescence.With increasing concentrations of quantum dots,the fluorescence intensities representing the levels of OCT4 and Nanog in iPSCs were gradually elevated,with optimal levels of fluorescence observed at a concentration of 10 nmol/L of quantum dots.The fluorescent labeling of OCT4 and Nanog in iPSCs remained and weakened gradually till day 7 even after the second passage.Conclusions Quantum dots labeling could be used to track iPSCs in a dose-independent manner.The fluorescent signal from the quantum dots labeling the iPSCs lasts 2 weeks at least.

Objective:To compare and explore the effects of needling acupoints at different nerve segmentson the oxytocin (OT) neurons in the paraventricular nucleus of hypothalamus (PVN) and the intragastric pressure, and discuss the possible mechanisms. Methods: Thirty-two healthy adult Sprague-Dawley (SD) rats were numbered and divided into 4 groups according to the random number table, a Zusanli (ST 36) group, a Neiguan (PC 6) group, a Weishu (BL 21) group and a control group, with 8 rats in each group. Except the control group, rats in the other three groups received acupuncture at the corresponding acupoints. To observe the differences in double-labeled OT neurons and c-fos neurons of the hypothalamic PVN and the intragastric pressure after acupuncture among the three groups of needling acupoints at different nerve segments. Results:Compared with the control group, the numbers of double-labeled cells in the PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, while the intragastric pressure increased significantly (allP<0.05), and the inter-group differences were statistically significant (P<0.05). The intragastric pressure in the Weishu (BL 21) group decreased significantly, and the inter-group difference was statistically significant (P<0.05). Compared with the Weishu (BL 21) group, the numbers of OT/c-fos double-labeled cells in PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, and the intragastric pressure increased significantly, the inter-group differences were statistically significant (allP<0.01). Conclusion:Acupoints at different nerve segments have different regulation effects on intragastric pressure. The difference may be related to the different nerve conduction pathways by acupoints at different nerve segments in regulating the intragastric pressure. The PVN may be one common integration center for the regulation of gastric function in the three acupoints [Zusanli (ST 36), Neiguan (PC 6) and Weishu (BL 21)] at different nerve segments.

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 study the correlation between level of nitric oxide/ nitricoxide synthase(NO/NOS) on placenta homogenate and ultra-structure changes of placenta in pregnancy lead exposure in rats.Methods Seventeen normal pregnant rats and 46 rats of exposured in lead which were divided into A,B,C groups were studied.The level of NO/NOS of placenta were measured by nitrate reductase and NOS kit.Placentas were randomly selected from each group to detect ultra-structure by electron-microscope.Results There were significant difference among A,B and control groups on level of NO/NOS(all P0.05).Compensation hyperplasy or minor injury were observed in lead exposure of stage groups.Lead exposure during whole gestation period,the lead level was maxmum,and decompensation were observed on placental construction.Conclusions There is a close correlations between level of lead,NO/NOS and pathological change of placental tissue,and both of them may play an important role in the pathogenesis of peripartum lead exposure.

BACKGROUND: Magnalium which is potential to be the medical biodegraded metal implant is more and more interesting,but it must be well biocompatibility to human body.OBJECTIVE: To evaluate the sensitization of magnalium (AZ31B).METHODS: A total of 35 guinea pigs were randomly divided into saline group (negative control group,n=10),5% volume of formaldehyde (positive control group,n=10),and AZ31B group (n=15).Sensitization test at the maximal dosage was performed according to "Biological evaluation of medical devices-Part 10: Tests for irritation and delayed-type hypersensitivity",including intracutaneous induction,local induction,and provocation.Patch was removed after 6,24,48,and 72 hours,and the skin response was classified accordingto Magnusson and Kligman criteria.Patch was removed after 72 hours,and skin was performed with biopsy,stained with FIE staining,and observed under optic microscope.RESULTS AND CONCLUSION: Sensitization response was not tested in both negative control group and AZ31B group at 24,48,and 72 hours after patch removal; however,moderate erythema was observed in the positive control group.Optic microscope demonstrated that criteria of allergy such as spongiosis,edema,and diffuse as well as perivascular mononuclear infiltration was not observed in the AZ31B group,but a few basophilic calls ware observed.This suggested that AZ31B was biologically safe for sensitization.