Background Transforming growth factor-β1(TGF-β1) plays an important role in corneal woundhealing.The effects of TGF-β1 on the synthesis of extra cellular matrix (ECM) vary upon different concentrations.Previous studies focused on the effects of high concentration of TGF-β1 on keratocytes under the two-dimensional culture condition,and the effect of low concentration of TGF-β1 on the synthesis of ECM in keratocytes remains unclear.Objective This study was to investigate the growth of Pellet,a three-dimensional model of corneal stroma cells in vitro,and its ECM synthesis under a low concentration of TGF-β1.Methods Bovine corneal stromal cells were isolated from fresh bovine eyeballs by two-step digestion by collagenase and cultured using DMEM/F12 medium with 10％ fetal bovine serum (FBS).Pellets derived fresh bovine keratocytes with culture medium containing 0.25 ng/ml TGF-β1 +5％ FBS and 0.50 ng/ml TGF-β1 +5％ FBS were established,respectively.The morphology of Pellets was observed under the natural light at 48 hours,1 week,2 weeks and 3 weeks after culture.In 3 weeks after culture,the cell structures was observed by hematoxylin-eosin staining,and Calcein-AM/propidium (Calcein-AM/PI) staining was used to assay the cell viability.Real-time flurorescence quantitative PCR and immunofluorescence technology were applied to analyze the expressions of α-smooth muscle actin (α-SMA),fibronectin (FN),type Ⅰ collagen (Col Ⅰ) and type Ⅲ collagen (Col Ⅲ) mRNA and proteins.RT-PCR was employed to detect the expressions of lumican (LUM) mRNA and keratocan (KERA) mRNA in the cells.Results Cells in Pellet clustered throughout the culture duration.Hematoxylin-eosin staining showed the mass red-dyed collagen fibers in both 0.25 ng/ml TGF-β1 +5％ FBS group and 0.50 ng/ml TGF-β1 +5％ FBS group,and most cells possessed complete structures.The death rate of the cells was (33.60±1.65)％ in the 0.25 ng/ml TGF-β1 +5％ FBS group and (30.90±0.78) ％ in the 0.50 ng/ml TGF-β1 +5％ FBS group,showing an insignificant difference between them (t =0.144,P=0.887).The expressions of α-SMA,FN and Col Ⅲ proteins in 0.25 ng/ml TGF-β1 +5％ FBS group were lower than those in the 0.50 ng/ml TGF-β1 + 5 ％ FBS group (tα-SMA =4.622,P =0.010;tFN =2.973,P =0.040;tCol Ⅲ =7.845,P＜0.001),but the expression of Col Ⅰ in 0.25 ng/ml TGF-β1 +5％ FBS group was higher than that in 0.50 ng/ml TGF-β1+5％ FBS group (tColⅠ =4.022,P=0.016).The ratio of Col Ⅲ/Col Ⅰ in 0.25 ng/ml TGF-β1+5％ FBS group was lower than that in the 0.50 ng/ml TGF-β1 +5％ FBS group in both mRNA and protein level (tmRNA =-3.039,P =0.038;tprotein =3.215,P =0.032).The expression of LUM mRNA and KERA mRNA were detected in Pellet at different time points.The expression of LUM mRNA in 0.25 ng/ml TGF-β1 +5％ FBS group increased over time.While in 0.50 ng/ml TGF-β1 +5％ FBS group,the expression of LUM mRNA peaked at 1 week but declined at 2 weeks.The expression of KERA mRNA in two groups were all peaked at 1 week but declined at 2 weeks.Conclusions Low-dose TGF-β1 in Pellet can maintain the normal growth of keratocytes and synthesize ECM.The expression of ECM tends to the normal condition after reducing the concentration of TGF-β1,implying a scarless expression.

Leukoaraiosis (LA) has important clinical significance;however, the neurobiological mechanism was still unclear. In this pa-per, we reviewed literatures about the clinical symptoms, pathology and imaging of LA in order to improve the understanding of the patho-logic mechanism, recognize the reversible pathological change in the earliest stage, which can help to prevent dementia.

Objective To explore the related risk factors of cerebral hemorrhage complicated with stress ulcer (SU). Methods The clinical data of 1 185 patients with cerebral hemorrhage admitted to Department of Emergency Medicine of Nanjing General Hospital from March 2006 to March 2014 were retrospectively analyzed. Patients were divided into two groups according to whether patients complicated with SU or not. Data was collected within 8 hours after admission in two groups including gender,age,amount of bleeding,the bleeding site (basal ganglia,thalamus, brainstem,brain lobe,ventricle,subarachnoid,and cerebellum),disturbance of consciousness,acute physiology and chronic health evaluationⅡ(APACHEⅡ)score,systolic blood pressure(SBP),history of hypertension,and history of cerebral hemorrhage. The statistically significant risk factors found using univariate analysis was selected and was analyzed to find independent risk factors with multivariate logistic regression analysis. The receiver operating characteristic curve (ROC curve)was plotted to analyze the independent risk factors and evaluate their power of test. Results 1 185 patients with cerebral hemorrhage were enrolled in the study,293 cases occurred SU,accounting for 24.7%,and 892 cases without SU,which accounted for 75.3%. As shown by univariate analysis,risk factors for cerebral hemorrhage complicated with SU included age,amount of bleeding,the bleeding site,disturbance of consciousness,APACHEⅡscore,SBP. As to the site of bleeding,brain,thalamus,brainstem hemorrhage complicated with SU were higher proportion,45.3%(43/95),39.1%(63/161),36.9%(48/130),which were significantly higher than those of the lobes of the brain 〔26.2% (33/126)〕,cerebellum 〔18.8% (15/80)〕,basal ganglia〔16.1%(78/485)〕,arachnoid the inferior vena cava 〔12.0% (13/108)〕. Multivariate logistic regression analysis showed that amount of bleeding 〔odds ratio (OR)=3.305,P=0.001,95%confidence interval (95%CI)2.213-48.634〕,the bleeding site (OR=1.762,P=0.008,95%CI 0.123-2.743),SBP (OR=1.223,P=0.034,95%CI 0.245-2.812) were independent risk factors of cerebral hemorrhage complicated with SU. The area under the ROC curve (AUC)of amount of bleeding and SBP were 0.846 and 0.597,suggesting that amount of bleeding has moderate diagnostic value and SBP has low diagnostic value. Conclusions Cerebral hemorrhage patients with large amount of bleeding,the bleeding site in the ventricle,thalamus or brainstem,high SBP are of great risk. We should lower blood pressure and give preventive treatment for SU as soon as possible.

Objective:To determine the half lethal dose(LD50) of paraquat in rats and to establish a relatively safe and stable pathological animals model of pulmonary fibrosis.Methods: Ninety-six SD rats totally in half genders.Fifty SD rats in half genders were randomly divided into 5 groups,each had 10 rats.Feed the rats with different doses of liquor of paraquat intraperitoneally one time and definite the half lethal dose of one and two weeks.After that,prepare another forty-six SD rats,also in half genders,as intonication group,twenty-eight rats were treated with the liquor of paraquat in dosage of 18 mg/kg intraperitoneally.As control group,sixteen rats were treated with equivalent volume of normal saline.Observe the toxic symptom daily and rats were sacrificed on day 1,3,5,7,14,21,28,35 and 42 respectively for the histological examination.Results: The half lethal doses of intraperitoneal paraquat of 1 and 2 weeks were 18.27 and 17.29,with 95% confidence intervals of 16.61-20.09 and 15.99-18.67,respectively.After intraperitoneal paraquat injection at the dose of 18 mg/kg,typical toxic symptoms were observed at different times in the rats.The whole process of acute lung injury and fibrosis induced by paraquat intoxication could be seen with the naked eyes or under the light microscope.Conclusion: Paraquat has a strong toxicity to rats.A proper dose of paraquat solution can not only reduce the number of experimental rats,but also induce typical pulmonary fibrosis in rats.

Objectives:To investigate the risk factors for acute liver injury (ALI) in patients after resuscitation from cardiac arrest and their influence on prognosis.Methods:The clinical data of patients after cardiopulmonary resuscitation in our department from January 2015 to January 2018 were analyzed. According to whether ALI occurred, the selected patients were divided into the ALI group and non-ALI group. The basic situation of the two groups of patients and the occurrence of shock and cardiac insufficiency after cardiac arrest were investigated. Kaplan-Meier method was used to analyze the effect of ALI on the 1-year survival of patients. The 28-day mortality and neurological recovery were observed in patients in the ALI group. Multivariate logistic regression was used to analyze the risk factors for ALI.Results::There were 54 patients in the ALI group and 158 patients in the non-ALI group. The patients in the ALI group needed a longer time to recover spontaneous circulation [19 (10-27) min, P=0.015], and the overall condition (SOFA score, acidosis, and lactic acid) were more serious. The incidences of shock and heart failure after cardiac arrest in the ALI and non-ALI groups were 74% and 55%, and 89% and 70%, respectively. The 1-year cumulative survival rate of patients in the non-ALI group was significantly higher than that of the ALI group ( P=0.043). The longer the duration of ALI, the higher the incidence of poor prognosis. The time to resume spontaneous circulation ( OR=3.762; 95% CI: 2.347-5.098) and heart failure ( OR=4.272; 95% CI: 2.943-5.932) after cardiac arrest were associated with ALI in patients after cardiopulmonary resuscitation (both P<0.05). Conclusions:The time to resume spontaneous circulation after cardiac arrest and heart failure after cardiopulmonary resuscitation are risk factors for ALI, and the occurrence of ALI increases patient’s mortality.

Objective Acute coronary syndrome ( ACS) is frequently accompanied by chronic comorbidities , which may se-riously affect its prognosis .This study aims to investigate the value of the Charlson Comorbidity Index ( CCI) in predicting the outcome of ACS by assessing the impact of individual and post-weighted-assignment comorbid conditions of the disease . Methods We retro-spectively analyzed the clinical data on 1 096 cases of ACS treated in Jinling Hospital from January 2010 to March 2014 .We reviewed their general information , clinical presentations , complications , and previous treatments , calculated CCI , and used in-hospital mortali-ty as the index for judging the prognosis . Results Of the 1 096 patients, 73%were males (aged 64.2 ±12.9 years), 27% were females (aged 72.1 ±12.6 years), and 46.8% had comorbidities. Of the diseases included in the CCI system , previous myocardial infarction was the most frequent comorbidity (18.0%), followed by diabetes mellitus ( 14.7%), moderately to severe renal disease (7.1%), cerebrovascular disease (6.0%), and chronic lung dis-ease (6.0%).Single factor analysis revealed statistically significant differences between different CCI groups in such clinical indicators as history of coronary artery disease , history of hypertension , time between symptom onset and admission , hemodynamics , drugs adminis-tered (aspirin, P2Y12 blockers, ACEI/ARB or statins), and reperfusion therapy (P<0.05).Logistic regression analysis showed the strongest predictors of in-hospital mortality were heart failure (OR 1.88, 95%CI:1.57-2.25), metastatic tumor (OR 2.25, 95%CI:1.60-3.19), renal disease (OR 1.84, 95% CI:1.60-2.11), and diabetes mellitus (OR 1.35, 95% CI:1.19-1.19). Receiver operating characteristic curve analysis manifested that either CCI with age or CCI with age and gender was superior to CCI a -lone in predicting in-hospital mortality of ACS patients (AUC 0.761 [95%CI 0.748-0.773] and 0.756 [95%CI:0.743-0.768] vs 0.670 [95%CI:0.656-0.685]). Conclusion Heart failure, diabetes mellitus, renal disease, and metastatic tumors contrib-ute to the in-hospital mortality of ACS patients .CCI together with age and gender may help to assess the prognosis of the disease .

OBJECTIVETo determine the feasibility of water fluoridation to prevent caries in Kunming by investigating the epidemiological status of dental caries and dental fluorosis of children, and to provide the longitudinal reference data for the long-term epidemiology survey of dental caries and dental fluorosis in Kunming city.

METHODSThrough stratified cluster sampling method, 212 5-year-old children and 1149 12-year-old children were recruited in the survey. Dental caries condition of each child was clinically examined, dental fluorosis was examined in 12-year-old group.

RESULTSThe prevalence of dental caries of primary teeth in 5-year-old group was 73.6%, mean value was 4.47 +/- 4.39. The values of permanent teeth in 12-year-old group were 53.5% and 1.42 +/- 1.83. The prevalence of dental fluorosis in 12-year-old group was 4.1% and the average community fluorosis index was 0.03.

CONCLUSIONBased on the high prevalence of dental caries and the low prevalence of dental fluorosis, it is suggested that using water fluoridation to prevent caries is feasible and necessary in Kunming city.

Objective To investigate the clinical value of the simple clinical score (SCS) in Emergency Department.Methods A total of 655 patients with critically illness admitted from July 1,2011through August 31,2011 were enrolled to evaluate the clinical application of SCS in emergency rescue room by analysis of the relationship between SCS and outcome of patients,and the correlation between each factor of SCS and the risk of death. Results The higher SCS,the higher hazard ratio for death.Differences in the hazard ratio for death among groups with different scores of SCS were statistical significance ( P ＜ 0.01 ).According to Chi-square test,there were significant differences in each factor ( except body temperature &unable standing or need Home Health Aide) among the groups of SCS ( P ＜ 0.05 ). Binary logistic regression analysis of each factor showed that age,diabetes,coma,pulse,systolic pressure and respiratory rate had significant correlation with patient mortality. After logistic regression analysis,age,diabetes,coma,pulse,systolic pressure and respiratory rate are significantly related to the mortality of patients.Conclusions The SCS scoring system is useful to make a precise evaluation of critically ill patients in the emergency department.When emergency rescue is carried out,particular stress should be focused on age,diabetes,coma,pulse,systolic pressure and respiratory rate.

Objective Though paraquat (PQ) is highly toxic, there is still no effective treatment for PQ poisoning .The aim of the article was to study the protective effect and mechanism of the p 38 mitogen-activated protein kinase ( MAPK) inhibitor SB203580 on PQ-induced acute lung injury in rats . Methods 72 SD rats were randomly divided into three groups ( n=24 ): normal saline (NS) group, PQ poisoning group and p38 inhibitor SB203580 intervention (PQ+SB) group.The arterial blood gas analysis, lung wet and dry ratio (W/D),the expression of tumor necrosis factor-α(TNF-α), the superoxide dismutase (SOD) level and the pathological changes of lung tissues were recorded at different time points after drug intervention . Results On the 1st , 3rd, 5th days after drug intervention in PQ group, the alveolo-arterial oxygen partial pressure difference (PA-aO2) [(45.67 ±4.17), (68.78 ±6.63), (80.23 ±7.12 ) mmHg ], the lung tissue TNF-αexpression (14.63 ±3.10], [18.24 ±2.98], [16.22 ±2.79] pg/mg) and W/D ([4.931 ±0.034], [5.020 ±0.064], [5.079 ±0.016]) in-creased gradually to a peak on the 3rd day, while the SOD level de-creased respectively on the 1st , 3rd, 5th days after drug intervention ([175.26 ±7.98], [167.57 ±8.05], [160.24 ±6.78] U/ug) (P<0.05).Compared with PQ group, PQ+SB group got a decrease in the PA-aO2([80.23 ±7.12] vs [44.17 ±4.16]), the lung tissue TNF-αexpression ([16.22 ±2.79] vs [9.48 ±2.72]) and W/D ([4.805 ±0.070] vs [5.079 ±0.016]) (P<0.05), while the pulmonary SOD level increased in comparison with PQ group ([125.89 ±6.65] vs [160.24 ±6.78]) (P<0.05). Conclusion The p38MAPK inhibitor SB203580 plays a certain protective role in PQ-induced acute lung injury by reducing inflammation and improving antioxidant capacity .

Objective To explore the effects of intestinal trefoil factor ( ITF) on gastric mucosal epithelial cell proliferation and its possible molecular mechanism . Methods The cultured GES-1 cells were treated with ITF in the concentration of 100 ng/mL and 500 ng/mL in vitro, and then were observed using microscope for the morphological analysis .The Cell Counting Kit-8 ( CCK-8) was used to detect the proliferation activity of GES-1.The cultured GES-1 cells were treated with 100 ng/mL ITF and the specific inhibitor of PI3K/Akt signaling pathway LY294002 (15μmol/L) in vitro, and then were observed using microscope for the morphological analysis . The proliferation activity of treated GES-1cells was detected using CCK-8 and the expressions of p-Akt and Akt of PI3K/Akt signaling pathway were determined by Western blot . Results Compared with the control group , the proliferation activity of GES-1 cells in-creased after being treated with ITF and the higher concentration of ITF induced the higher proliferation activity .LY294002 inhibited the increased proliferation activity of GES-1induced by ITF.The data of Western blot indicated that ITF induced the expression of p -Akt and activated the P3IK/Akt signaling pathway to modulate the proliferation activity of GES -1 cells.However, LY294002 inhibited the PI3K/Akt signaling pathway and then decreased the proliferation activity of GES -1 cells. Conclusion ITF could promote the proliferation ac-tivity of GES-1 cells by activating PI3K/Akt signaling pathway .