Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Medicine, Chinese Traditional ; methods ; Systems Biology

Animals ; Astragalus Plant ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Liver Cirrhosis ; drug therapy ; Phytotherapy ; Pulmonary Fibrosis ; drug therapy ; Rheum ; Salvia miltiorrhiza ; chemistry

Objective Through the analysis of cases,to improve the cognition of clinicians on extracranial vas-cular embolism diseases(VED)in children.Methods The clinical information included incidence trend,thrombophi-lia,clinical features,treatment and prognosis of 29 children with VED,who were admitted from January 201 1 to Decem-ber 201 5 in Nanjing Children′s Hospital Affiliated to Nanjing Medical University,and retrospectively analyzed.Results The annual rate of VED increased from 0 to 1 .49 per 1 0 000 hospital admissions from January 201 1 to December 201 5.In 29 cases,there were 20 male and 9 female,with an average age of 6.03 years old (2 h -1 3.83 years old).In-fants accounted for >20%(6 /29 cases).The average age of the female,non infected group,rheumatic disease group and venous thrombosis group was higher than that of the male,the infection group,the non rheumatic disease group and the arterial embolization group,but there was no significant difference(all P >0.05).Thrombophilia were as follows:in-fection disease,heart disease,kidney disease syndrome,rheumatoid disease,tumor,and twins embolization syndrome. Risk factors included central venous line,immobility,dehydration,glucocorticoids use,etc.Heart disease was the leading cause of non thrombotic embolism,while other thrombophilic diseases resulted in thrombotic embolism.Pulmonary em-bolism was mainly seen in patients with severe pneumonia,especially mycoplasma pneumoniae pneumonia (MPP ).The older the age,the more typical clinical manifestations,and significantly elevated D dimer,fibrin/fibrinogen degradation products tips VED.Treatment depends on the degree of risk.Thirty -seven point five percent(3 /8 cases)of children with arterial embolism were less than 1 year old,and the mortality 25.00% (2 /8 cases)was significantly higher than that of venous thrombosis 1 7.65%(3 /1 7 cases)and 1 1 .76% (2 /1 7 cases).Conclusion Heart disease mainly cause arterial embolism and infection,while nephritic syndrome and rheumatic disease mainly cause venous embolism.Pulmona-ry embolism is mainly seen in children with severe pneumonia,especially MPP.The risk of arterial embolization is higher than that of venous embolism.

Objective To compare the efficacy of internal fixation with non-operative treatment of traumatic flail chest combined with sternal fracture.Methods A retrospective review was made on 102 patients with traumatic flail chest combined with sternal fracture treated from January 2007 to January 2014.There were 71 patients in operation group and 31 patients in non-operation group.Operation group was allocated to pure titanium rib plating (Group Ⅰ,22 cases),porous titanium screw fixation (Group Ⅱ,20 cases),and nickel-titanium memory alloy sternal rib plating (Group Ⅲ,29 cases).Rib traction was performed in non-operation group.Clinical results and incidence of complications were compared between the two groups.Results At 24 and 72 hours,operation group was superior over non-operation group with regard to HR [(100.4 ± 9.5) times/min vs (105.2 ± 10.1) times/min,(92.1 ± 9.9) times/min vs (102.1 ± 9.2) times/min],MAP [(97.7 ± 14.5) mmHg vs (91.0 ± 13.3) mmHg,(112.5 ± 15.2) mmHg vs (93.1 ± 13.8)mmHg] and CVP [(7.8 ±3.7)cmH2O vs (9.4 ±3.8)cmH2O,(6.2 ±2.9)cmH2O vs (9.1 ±3.2) cmH2 O] (P ＜ 0.05).Duration in ICU [(5.1 ± 0.8) days vs (9.3 ± 1.1) days],ventilation time [(4.5 ±1.0)days vs (8.2 ± 1.4)days],hospitalization time [(14.6 ±3.5)days vs (23.3 ±4.4)days] and incidence of complications were significantly lower in operation group than in non-operation group (P ＜ 0.05).Two months after discharge,tidal volume (VT) [(0.52 ±0.04)L vs (0.40 ±0.03)L],inspiratory capacity (IC)[(1.99 ±0.45)L vs (1.22 ±±0.33)L],forced vital capacity (FVC) [(3.52 ±0.51)L vs (2.44 ±0.42)L],total lung capacity (TLC) [(5.41 ±0.82)L vs (3.72 ±0.56)L] and forced expiratory volume in one second (FEV1) [(2.80 ± 0.43) L vs (1.95 ± 0.50) L] showed significantly better results in operation group than in non-operation group (P ＜ 0.05).Operation time,intraoperative bleeding and daily drainage volume were (38.8 ±9.2)min,(43.5 ±7.6)ml and (10.9 ± 1.1)ml in Group Ⅱ versus (62.5 ± 10.1)min,(100.0 ± 10.5) ml and (26.8 ±3.1)ml in Group Ⅰ and (49.3 ±9.6)min,(61.4 ±8.3)min and (19.8 ±1.6)ml in Group Ⅲ (P ＜ 0.01).Conclusion For traumatic flail chest combined with sternal fracture,the internal fixation should be done as early as possible so as to reduce incidence of complications.Relatively,porous titanium screw fixation has better clinical outcome.

Hip Joint ; Humans ; Ischium/diagnostic imaging* ; Magnetic Resonance Imaging

Objective To observe the expression of matrix metalloproteinases(MMP)-1,MMP-3 and iNOS in cartilage of experimental rabbit osteoarthritis at different time intervals after anterior cruciate ligament transection(ACLT)operation.The aim of this study is to provide the theoritical evidence for using ACLT rabbit model in Osteoarthritis(OA)research.Methods Unilateral ACLT was performed on 27 randomly selected while rabbits and underwent unilateral arthrotomy was performed on the other 9 white rabbits as the control group.Nine randomly selected white rabbits in experimental group were killed and 3 white rabbits in the control group at 4th,8th and 12th week respectively.Cartilage degradation of femoral condyles was evaluated macr-oscopically,mRNA expression level and protein expression level of MMP-1,MMP-3 and iNOS was measured by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry respectively.Results Forepart OA cartilage degradation was observed at the 4th week and became more severe at the 8th week after ACLF.Afterpart cartilage degradation was evident at the 12th week after ACLT while cartilage still remained normal in the control group,mRNA expression level and protein expression level of MMP-1.MMP-3 and iNOS were increased at the 4th week and became higher gradually at the 8th,12th week after ACLT compared with the control group.Expression distribution of MMP-1,MMP-3 and iNOS bad different patterns respectively.Conclusion It is suggested that the process of OA cartilage degradation can be simulated by ACLT model and MMP-1,MMP-3 and iNOS may be good markers in therapeutical research of OA.

Objective To investigate the computed tomography (CT) features of autoimmune pancreatitis (AIP).Methods The CT imaging data of 33 patients with AIP confirmed by pathology and/or steroid therapy were retrospectively analyzed.Image analysis including the shape of pancreas, density of lesion, contrast enhancement, the changes of pancreatic duct and biliary duct, peripancreatic appearances and adjacent organ involvement.T test was performed for statistical analysis.Results Among 33 patients with AIP, 23 cases (70%) with pancreatic parenchyma diffuse enlargement, eight cases (24%) with partial enlargement and two cases (6%) with normal pancreas.The lesions appeared hypoattenuating or isoattenuating on plain CT scan.After contrast-enhanced scan, the average CT values of lesions were (75.7±17.0) Hu at arterial phase, which was lower than that of venous phase (90.7±12.0) Hu, and the difference was statistically significant (t=3.378,P=0.002).The lesions demonstrated as progressive enhancement at venous phase.Among 33 patients, the main pancreatic duct was visible in six patients (18%).Sixteen patients (48%) presented with intrahepatic and extrahepatic biliary tract dilatation caused by intrapancreatic common bile duct stenosis.Thickened envelope-like structure around the lesions, presenting as capsule sign was seen in 14 patients (42%).Extra-pancreatic organ involvement was found in seven patients including three cases of kidney involvement.After treated with steroid, seven patients repeated CT which showed different degrees of improvement.Conclusion The main CT findings of AIP are diffuse and partial enlargement of pancreas with progressive enhancement at venous phase, envelope-like structure around pancreas, and stenosis of intrapancreatic common bile duct, which are important in the diagnosis and differential diagnosis of AIP.

Objective To investigate the expression and function of retinoic acid-inducible gene Ⅰ(RIG-Ⅰ) in monocyte-derived dendritic cells (MoDC) at different stages of hepatitis B virus(HBV)infection and to explore the role of RIG-Ⅰ in the disease progression after HBV infection. Methods Peripheral blood samples were collected from 28 hepatitis B virus-infected persons, including 21 cases of chronic hepatitis B (CHB) and 7 of acute hepatitis B (AHB). Eighteen healthy subjects were recruited as controls. Purified CD14~+ monocytes were isolated by CD14 microbeads. MoDCs were induced from CD14~+ monocytes with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4 for 7 days, and then were infected with vesicular stomatitis virus (VSV) to stimulate RIG-Ⅰ expression. The mRNA expression levels of RIG-Ⅰ, interferon (IFN )-promoter stimulating factor-1 (IPS-1) and IFN-β at 16 hours and 24 hours after infection with VSV were measured by real-time quantitative polymerase chain reaction (PCR). Data with normal distribution were tested by analysis of variance. Continuous variables between groups were compared using Mann-Whitney U test. Comparison among multiple groups was done by Kruskal-Wallis test. Results The expression levels of RIG-Ⅰ in MoDCs from CHB patients were significantly lower than those in AHB patients and healthy controls at 16 hours (2.44±2.03, 19. 54±3. 15, 21. 48±8. 39, respectively; F=7.451,P=0.002) and 24 hours (2. 68±2. 93, 10. 31 ±3. 88, 14. 01 ±5. 04, respectively, F = 7. 908, P = 0. 001)following VSV stimulation. The IPS-1 levels in both CHB patients and AHB patients were higher than those in healthy controls at 16 hours (2. 05±l. 08, 1. 99±1. 56, 0. 60±0. 31, respectively) F=7.246,P =0.003) and 24 hours (2. 27±2. 16, 3.24 ± 1.21, 1. 08±0. 73, respectively; F= 13. 598, P = 0. 001).Furthermore, the IFN-β expression levels were significantly lower in CHB patients compared to AHB patients and healthy controls at 16 hours and 24 hours after VSV stimulation. Conclusions The expressions of RIG-Ⅰ and IPS-1 in MoDC are abnormal in HBV infected persons, which indicates that RIG-Ⅰ signaling pathway might be blocked by HBV. The impaired function of MoDC may play a role in HBV infection and chronicity.

0.05).Conclusion Compared with multi-breath hold GRAPPA technique,single breath hold TSENSE technique is a fast,accurate and reproducible method and can be an substitute for GRAPPA to evaluate ventricular function.