Objective:To analyze the different of water fasting time on fiberoptic bronchoscopy examination. Methods: Chosen patients in our hospital received fiberoptic bronchoscopy examination as research subjects, randomly divided into control group with 4-6 h preoperative fasting and observation group with 2 h fasting, and compared blood glucose levels, stress levels, depression and anxiety score. Results: 1) observation group patients’ blood glucose values after water fasting were significantly higher than control group patients (t=5.382, t=5.893, t=6.624, t=7.934; P<0.05);2)observation group patients’ stress levels were significantly lower than control group patients (t=4.826, t=7.263, t=6.095; P<0.05); 3) observation group patients’ depression and anxiety scores before fasting had no difference(t=0.637, t=4.958; P>0.05), SAS and SDS scores of observation group patients 1h before the examination and inspection were significantly lower than the control group patients (t=6.382, t=6.562, t=4.958, t=7.632;P<0.05). Conclusion:Two hours before bronchoscopy is more conducive to the maintenance of water fasting blood sugar stable patients, reduce stress levels, relieve negative emotions in patients with positive clinical significance.

Objective To identify the promoter sequence of endothelial-overexpressed lipopolysaccharideassociated factor 1 ( EOLA1) gene and to elucidate the molecular mechanisms controlling EOLA1 expression. Methods A DNA fragment containing 1 723 bp 5' upstream of the EOLA1 gene and the transcription start site was generated by polymerase chain reaction and then cloned into a luciferase reporter gene vector,pGL3-basic. The relative luciferase activities driven by this 5'-upstream fragment and a series of deletion mutants were measured in transiently transfected human ECV304 cells,respectively. At last,the 1 723 bp upstream of the EOLA1 gene was analyzed online with Cluster Buster. Results A fragment 785 bp upstream of the EOLA1 coding region was sufficient to promote transcription. Further deletion analysis of the 785 bp fragment indicated that a 68 bp element from-738 to -676 was important for EOLA1 transcription in ECV304 cells. The 1 723 bp sequence contains binding sites for Sp1 and Myf. Conclusion We map the EOLA1 promoter by deletion analysis and reveal that the proximal region ( -738 to -676 bp) ,which contains binding sites for Sp1 and Myf,is essential for human EOLA1 promoter activity in ECV304 cells.

Aim To observe the effect of mycophenolate mofetil(MMF) on expressions of MCP-1 and CD68 in renal tubulointerstitial injury of diabetic rats and explore the mechanism of MMF′s protective role.Methods Diabetes was induced in uninephrectomized male Wistar rats by peritoneal injection of STZ (65 mg?kg-1). Rats were randomly divided into three groups:control group (NC),diabetic group (DM) and treated group (DM+MMF) with MMF(15 mg?kg-1?d-1).This study lasted for 8 weeks. 24 h urinary protein,blood glucose and the ratio of left kidney weight/body weight were determined after 8 weeks.The renal tubulointerstitial morphological change was observed,immunohistochemical method was used to analyze expressions of MCP-1 protein and CD68. Expression of MCP-1 mRNA in renal tissue was measured by quantitative Real-time PCR.Results Compared with NC group, serum glucose level,24 hour urinary protein and the ratio of left kidney weight/body weight were significantly increased(P

Objective To discuss the effects of capsular tension ring after phacoemulsification combined with IOL implantation on tilt and decentration of IOL in high myopia patients with cataract by ultrasonic biomicroscope (UBM).Methods A total of 36 cases (40 eyes) with high myopia and cataract underwent phacoemulsification combined with IOL implantation were chosen.The average axial length was 26.88 mm.The patients were divided into implant group (19 eyes,the capsular tension ring was implanted) and control group (21 eyes,the routine surgery was performed).The patients were examined by conventional slit lamp,and the best corrected visual acuity (BCVA)was measured at pre-operation and postoperative 6 months.Tilt and decentration were measured horizontally and vertically,and total tilt and decentration were calculated by geometry method.Results The postoperative BCVA in the two groups were all better than the pre-operation,there was no statistical difference in the preoperative and postoperative BCVA between two groups (all P ＞ 0.05).The horizontal,vertical and total decentration the implant group were (0.15 ± 0.07) mm,(0.30 ± 0.40) mm,(0.11 ±0.02)mm,respectively,which in the control group were (0.26 ± 0.19)mm,(0.32 ±0.60) mm,(0.24 ± 0.97) mm,respectively.The horizontal,vertical and total tilt in the implant group were 0.02° ±0.11°,0.70° ±0.25°,0.21° ±0.74°,respectively,which in the control group were 0.11 ° ± 0.31 °,1.09° ± 0.20°,1.24° ± 0.97°,respectively.There were statistical differences in the horizontal,total tilt and decentration between two groups (all P ＜ O.05),but no statistical difference in the vertical tilt and decentration (P ＞ 0.05).Conclusion The capsular tension ring can stable the IOL position after surgery in high myopia and cataract patients.

Objective:To investigate the role of growth arrest specific protein 6 (Gas6) in regulating macrophage polarization in wound healing.Methods:Clean male B6 mice were randomly(random number) divided into the normal group, skin defect group, skin defect group + normal saline group (PBS group), skin defect + Gas6 (1 μg) group, skin defect + Gas6 (5 μg) group, and skin defect + Gas6 (10 μg) group. Ten mice in each group were used to observe the healing of skin wounds. Macrophages were isolated from the wound tissues of the remaining 6 mice on the fifth day after modeling. The levels of IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA), the mRNA expression levels of arginase-1 (Arg-1) and inducible nitric oxide synthase (iNOS) were detected by RT-PCR, and flow cytometry was used to detect the expression of M1 marker CD197, M2 marker CD163 and F4/80. HE staining was used to detect the pathological changes of skin wounds. Masson staining was used to analyze the granulation tissue and collagen deposition.Results:Scab began to form on the surface of the wound on the third day after the skin defect model was established. The wound area of the Gas6 treatment group was smaller than that of the PBS group, and the wound healing was better than that of the PBS group. Compared with the normal group, the proportion of CD197 in macrophages of the skin defect group was significantly increased ( P=0.00 49), the proportion of CD163 and F4/80 double positive was significantly decreased ( P=0.00 86), the level of IL-6 was significantly increased ( P=0.00 13), the level of IL-10 was significantly increased ( P=0.00 14), the level of iNOS mRNA was significantly increased ( P=0.00 8), and Arg-1 was significantly increased in the skin defect group The mRNA level was significantly decreased ( P=0.01 21), and the inflammatory infiltration was aggravated. Compared with the PBS group, the proportion of CD197 in the Gas6 treatment group was significantly decreased ( P=0.00 0), the double positive rates of CD163 and F4/80 were significantly increased ( P = 0.00 0), the level of IL-6 was significantly decreased (P = 0.00 0), the level of IL-10 was significantly increased ( P=0.00 03), the level of iNOS mRNA was significantly decreased ( P=0.00 18), the level of Arg-1 mRNA was significantly increased ( P=0.00 1), and the number of inflammatory cells and the number of collagen fibers were increased. Conclusions:Gas6 can promote the transformation of macrophages from M1 to M2 in mice with skin defect, which is beneficial to the wound healing of skin defect.

Objective To investigate the relationship between Klotho and autophagy in sepsisinduced acute kidney injury mice model.Methods The male healthy Balb/c mice were used to establish the model of sepsis-induced acute kidney injury by using cecal ligation and puncture (CLP).Mice were sacrificed at 3 h,6 h,12 h,1 d,2 d,3 d,and 5 d after CLP (n =12 for each interval) and on 1 d 6 mice in sham group as well as 6 mice in normal group were sacrificed at the same time.Scr and BUN in the blood serum were detected.The HE and PAS staining were employed for observation on the histopathological changes in kidney tissues under light microscope.The autophagosomes were observed under transmission electron microscope (TEM).The renal protein of Klotho,LC3 and P62 were detected by using Western blot and Immunohistochemistry.Statistical analyses were performed using Student's t-test by SPSS 23.0.software.Results Scr and BUN increased significantly after CLP,especially on 1 d,respectively (165.64 ± 20.56) μmol/L and (45.51 ± 4.05) mmol/L.HE and PAS staining showed renal tissue was damaged obviously 1 d after CLP,as indicated by desquamation of the brush border of proximal tubular epithelial cells,appearance of bare basement membrane,and interstitial inflammatory cell infiltration.Under TEM,autophagosomes and phagocytosis were observed.Compared with sham group,the expression of Klotho protein decreased gradually from 3 h to 1 d and dropped to the trough at 1 d (t =51.851,P ＜0.01),then resumed gradually from 2 d to 5 d.On the contrary,the activation of autophagy increased as indicated by the expression of LC3-Ⅱ/L3-Ⅰ and p62.Autophagy was induced gradually from 3 h to 1 d and reached peak at 1 d,then declined gradually from 2 d to 5 d (P ＜ 0.01).The protein of Klotho and LC3-Ⅱ mainly distributed in renal tubular cytoplasm,and Klotho was reduced significantly (t =-8.371,P ＜ 0.01) and LC3-Ⅱ appeared in high density remarkably (t =4.995,P =0.001) on 1 d after CLP.Conclusions Klotho protein reduction and autophagy protein increase were observed in sepsis-induced acute kidney injury,and the expressions of Klotho and autophagy acted out in certain extent of time dependence.

Adult ; Dinitrobenzenes ; poisoning ; Emergency Treatment ; Humans ; Male ; Middle Aged ; Occupational Exposure

Objective To examine the correlation of imbalance of urinary exosome Th1/Th2 with diabetic nephropathy (DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) and 30 healthy volunteers as control were enrolled in the study.According to urinary albumin/creatinine ratio (UACR),type 2 diabetes mellitus patients were divided into 3 groups:diabetes mellitus without nepbropathy group (DM,n=40,UACR＜30 mg/gCr),microalbuminuria group (DN 1,n=50,UACR-30～300 mg/gCr) and clinicoalbuminuria group (DN 2,n=30,UACR＞300 mg/gCr).Urine exosome-interferon-gamma (IFN-γ) and exosome-interleukin 4 (IL-4) levels were determined by enzyme-linked immunosorbent assay (ELISA).Multiple stepwise linear regression was used to analyze the correlation of exosome-IFN-γ/IL-4 with glycated hemoglobin (HbA1c),cholesterol (CH),UACR,Scr and BUN.Results Th1/Th2 ratio in DM,DN1,DN2 groups was significantly higher than that in healthy group (0.8089±0.2458,0.8993 ±0.3515,0.8571±0.2470 vs 0.6198±0.1769,all P＜0.01).Correlation analysis showed that urinary exosomeIFN-γ/IL-4 ratio was positively correlated with UACR (r=0.213,P=0.015) and BUN (r=0.292,P=0.001).Multiple stepwise linear regression analysis showed that BUN was independent determinants for exosome-IFN-γ/IL-4 (β=0.246,P=0.006).Conclusion The imbalance of urinary exosomeTh1/Th2 is correlated with DN,which may play an important role in the pathogenesis of DN.

Objective To discuss the protective effect of bone marrow mesenchymal stem cell(BMSC)on lung injury induced by vibrio vulnificus sepsis and its mechanism. Methods BMSCs were isolated by whole bone marrow adherent culture from mouse. Male ICR mice were randomly divided into normal saline control group(NS group),normal saline+BMSC control group(NSB group),vibrio vulnificus sepsis group(VV group),vibrio vulnificus sepsis + BMSC group(VVB group)according to random number table,with 40 mice in each group. Sepsis mouse model was reproduced by injecting vibrio vulnificus(1×107 cfu/mL)5 mL/kg through the left side peritoneal cavity, and caudal intravenous injection of BMSC(4×105 cfu/mL)5 mL/kg for intervention after model reproduction. Ten mice in each group were sacrificed at 6,12,24 or 48 hours after injecting vibiro vulnificus,and their lung tissues were harvested. The lung wet/dry(W/D)ratio was calculated. The expression of nuclear factor-κBp65(NF-κBp65)in nucleus was measured by Western Blot. The levels of tumor necrosis factor-α(TNF-α)and interleukins(IL-1β, IL-6)in lung tissue were detected by enzyme-linked immunosorbent assay(ELISA). The pathological changes in lung tissue were observed after hematoxylin-eosin(HE)staining and uranyl acetate-lead citrate staining. Results After vibrio vulnificus injection,lung W/D ratio,the expression of NF-κBp65 in nucleus,and the levels of TNF-α, IL-1β,IL-6 in the lung tissues were significantly increased in VV group compared with those in NS group at all the time points,and peaked at 12 hours. Compared with the VV group,the VVB group had significantly decreased levels of lung W/D ratio,NF-κBp65 expression,and the levels of TNF-α,IL-1β,IL-6,with significant differences at all the time points〔VV group vs. NS group at 12 hours:lung W/D ratio 7.22±0.03 vs. 5.21±0.02,NF-κBp65 expression (glay scale)1.86±0.74 vs. 0.75±0.07,TNF-α(ng/L)433.24±3.23 vs. 106.57±1.21,IL-1β(ng/L)35.64±0.15 vs. 10.64±0.48,IL-6(ng/L)58.84±0.55 vs. 17.69±1.35,all P<0.05;VVB group vs. VV group at 12 hours:lung W/D ratio 6.49±0.06 vs. 7.22±0.03,NF-κBp65 expression(A value)1.16±0.08 vs. 1.86±0.74,TNF-α(ng/L)357.22±3.25 vs. 433.24±3.23,IL-1β(ng/L)27.77±0.59 vs. 35.64±0.15,IL-6(ng/L)38.68±1.29 vs. 58.84±0.55,all P<0.05〕. There were no significant differences in above indexes between NS group and NSB group. In the NS and NSB groups pathological changes were not obvious under light microscopy,in the VV group lung tissue hyperemia and edema was significant,the edema fluid,red blood cells and inflammatory cells also could be seen, and in the VVB group lung damage that mentioned above could be alleviated. In the NS and NSB groups epithelial cell structure of type Ⅰ and type Ⅱ was completed,and the changes were not obvious under the transmission electron microscopy. In the VV group the alveolar walls were damaged significantly,with type Ⅰ epithelial cell cytoplasm swelling,bubbling and rupture,with type Ⅱ epithelial cells visible cytoplasm decrease,cavitation,addiction to osmium lamellar corpuscle emptying,lysosome hyperplasia,microvilli reduction,and in the VVB group the above damage was alleviated. Conclusion Vibrio vulnificus sepsis can cause acute lung damage and edema,and BMSC can down regulate inflammatory cytokines,reduce lung injury caused by vibrio vulnificus sepsis.

Objective: To investigate the effect of cycloartenyl ferulate (CF) on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells (HUVEC), and to explore its relative mechanism. Methods: Human umbilical vein endothelial cells were cultured in vitro, and the experiment was divided into the normal group, CF group, LPS group, and LPS+CF groups at different concentrations. After the corresponding treatment of cells in each group, the cell viability and apoptosis of each group were tested by the cell counter Kit-8 (CCK-8) assay and TdT-mediated dUTP nick end labeling (TUNEL) assay. The protein expression of Bax, Bcl-2, caspase-3 and the effect of CF on the protein expression of Nrf2/HO-1 pathway were determined by Western blot. One-way ANOVA were performed in multigroup mean comparison, LSD-t test was used to compare the mean values of two samples, and P<0.05 was considered statistically significant. Results: Compared with the LPS group, the survival rate of HUVECs cells in the CF group was significantly increased with different doses, and the survival rate increased significantly in the 320 μmol/L CF group [(69.85±1.2)% vs ( 100.2±1.824)%, P< 0.01]. TUNEL staining showed that compared with the LPS group, the number of apoptotic positive cells in the 320 μmol/L CF group was significantly reduced [(27.33 ± 3.40) vs (11.67 ± 2.04), P<0.01]. Compared with the LPS group, Bcl-2 protein expression level was significantly increased in the CF group at different doses (P<0.01), caspase-3 and Bax protein expression were significantly decreased (P<0.01). Nrf2 protein expression level increased significantly (P<0.01), and HO-1 protein level increased significantly (P<0.01). Conclusion CF can alleviate LPS-induced apoptosis of HUVEC, which may be related to the increase of bcl-2 expression, the decrease of caspase-3 and Bax protein expression and the activation of Nrf2/ HO-1 signaling pathway.