Objective To study the action and mechanism of staphylococcal exfoliative toxin A(E-TA)on pemphigus foliaceus antigen(PFA)and pemphigus vulgaris antigen(PVA)expressed on cultured human keratinocytes.Methods Stratified human keratinocytes were incubated with ETA and then stained with sera from patients with pemphigus foliaceus or pemphigus vulgaris as the first antibodies and FITC-la-beled sheep anti-human IgG as the second antibody.Total protein was harvested from the cells pretreated with ETA and run on SDS-PAGE for Western blot with the same antibodies.Simultaneously,supernatants of the keratinocytes before and after ETA treatment were collected for detection of the levels of IL-1?,IL-6with ELISA kits.The caseinolytic activities of the supernatants were tested by spectrometry in which casein was used as a non-specific substrate.Results Down-expression of PFA was shown after ETA treatment while no change of PVA expression was found.The high intensity and continuous linear appearance of fluo-rescent staining before ETA treatment became weak and discontinuous after ETA treatment,which were re-covered gradually in24hours.The degradation of proteins recognized by PF sera after ETV treatment was revealed by Western blot.The decreasing tendency of IL-1?concentration was found in the supernatants of cell culture after ETA treatment,but IL-6level was too low to be detected.Increased caseinolytic activities were found in the supernatants,and declined36hours after ETA treatment.Conclusions ETA acts on PFA expressed on keratinocytes in vitro,which is reversible along with withdrawal of ETA.The mechanism of E-TA act on PFA may be related to proteolytic action instead of promoting cytokine secretion.

Objective To investigate the effects of UVB irradiation and calciu m at different concentrations on the expression of pemphigus antigens by culture d human keratinocytes. Methods Human keratinocyte cultures were treated with eit her 2 mmol/L calcium added to the serum free media, or exposure to UVB irradiat ion. Immunofluorescence staining was performed with sera from patients with pemp higus vulgaris (PV) or pemphigus foliaceus (PF) as the first antigen. Extracts f rom both epidermis and keratinocytes were run on SDS-PAGE according to Laemmli ′s method and transferred to nitrocellulose membrane for immunoblot with PV and PF sera. Results Specific staining with PV sera was always detectable on kerati nocyte culture by immunofluorescence assay with or without high concentrations o f calcium while PF antigen was detected on stratified cells only. However, expos ure to UVB irradiation could not evoke keratinocyte culture express PF antigen. The reactivities were found at 160 kd band with PF sera while at both 160 kd and 130 kd bands with PV sera. Conclusions Monolayer or stratified keratinocytes ca n produce PV antigen, by increasing the concentration of calcium in the culture media, the human cultured kera tinocytes can be induced to stratify and express PFA. Human keratinocytes can not express PF antigen after exposure to UVB in intro.

Objective To compare the performances of baseline ultrasound(BUS),contrast-enhanced ultrasound (CEUS) and spiral computed tomography (CT) in characterizing of portal vein thrombosis (PVT).Methods The capability of differentiation of benign and malignant thrombi by using BUS,CEUS and spiral CT scan were assessed in 43 patients with PVT.Results The diagnostic accuracies of benign and malignant PVT were 100％ with CEUS,60.5％ with BUS and 76.7％ with CT,respectively.There were significant difference between every two groups( P ＜0.05).Conclusions CEUS is superior to BUS and CT in distinguishing benign and malignant PVT,thus it is recommended.

Objective To explore the change regulation of blood coagulation function before and after treatment of snake bite pa‐tients .Methods A total of 226 hospitalized patients with clear diagnosis belong to what kind of snakes in the past two years were selected in this study ,39 cases were bit by coral ,76 cases were bit by trimeresurus stejnegeri ,47 cases were bit by cobra ,24 cases bit by king cobra ,40 cases were bit by adder .Five blood coagulation indexes were determined before and after treatment various pe‐riods for these patients .The tested results were made to statistical analysis according to kind of snakes ,periods and disease condi‐tion .Results D‐D level obvious .rise before treatment for the severe cases patients of coral snake bite (P<0 .05) .Plasma prothrom‐bin time (PT) ,activated partial clotting enzyme live time (APTT) ,thrombin time (TT)and D‐D level rise and Fib level reduced be‐fore treatment for the severe cases patients of trimeresurus stejnegeri bite ,and Fib and D‐D level before and after treatment had sta‐tistical significant difference with the mild patients (P<0 .05) .Every index for the cobra and king cobra had no statistical signifi‐cant differences (P>0 .05) before and after treatment ,only D‐D level of the severe cases had statistical significance before treat‐ment in contrast to the mild cases of cobra bite (P< 0 .05) .The changes of 5 item blood coagulation indexes were all very large whether mild or severe for viper bite patients ,a number of comparisons had statistical significance difference (P<0 .05) whether before or after treatment and whether mild or severe for viper bite patients .Conclusion The bite of coral snake ,cobra and king co‐bra affect little for the blood coagulation function .The severe cases patients of trimeresurus stejnegeri bite might lead to more seri‐ous blood coagulation function abnormality but the recovery is faster after treatment .The viper bite might lead to maximum blood coagulation function abnormality and the recovery is slow after treatment .

Objective To determine the prevalence of antibodies to cyclic citrullinated peptides (antiCCP) in patients with juvenile-onset systemic lupus erythematosus (JSLE) and its potential clinical significance. Methods Anti-CCP was measured in sera from patients with JSLE (n=47), juvenile idiopathic arthritis (JIA, n=54) and the sera from age-matched healthy children (n=40) using the third generation of anti-CCP ELISA commercial kit. The association of anti-CCP with other laboratory parameters and clinical features, especially arthritic symptoms in JSLE was also analyzed. T-test, Mann-Whitney U test, Chi-square and Fisher's exact test were used for statistical analysis. Results Out of the 47 JSLE patients, 6 (13%) were anti-CCP positive, which was significantly higher than that of the healthy controls( 13% vs 0, P＜0.05 ), but not different from that of the JIA group (26%, P=0.098). RF was more prevalent in JSLE patients with anti-CCP than patients without (83% vs 15%, P＜0.01 ), but there was no difference in other laboratory parameters and the clinical features ineluding the occurrence of arthritis (67% vs 51%, P＞0.05). As one of the initial symptoms, arthritis was observed in 25 of 47 JSLE patients and no one had developed deforming arthropathy.There was no statistical difference in anti-CCP positivity between JSLE patients with and without articular involvement ( 16% vs 9%, P＞0.05 ). Anti-CCP was not detected in any of the 3 patients with JSLE who had experienced joint pain and limited activity during 3 years follow-up. Conclusion Anti-CCP could be detected in patients with JSLE. It is noteworthy when differentiate from juvenile idiopathic arthritis, but the presence of anti-CCP does not relate with the occurrence of arthritis at presentation and persistence of arthritis in JSLE.

Objective: To explore the correlation between the expression of C-terminal tensin-like protein (CTEN) and the prognosis of hepatocellular carcinoma (HCC). Methods: Using immunohistochemistry, we detected CTEN protein level in samples of primary lesion and adjacent non-tumor lesion collected from 240 patients with HCC. The relationship between CTEN expression and clinicopathology, 5 year recurrent-free survival, or overall survival was evaluated by Chi-square test, Kaplan-Meier, or Cox regression analysis. Results: High CTEN expression was detected in 55% of hepatocellular carcinoma tissues and 20%of adjacent carcinoma tissues (P<0.001). CTEN expression was positively correlated with tumor diameter (P=0.022), venous invasion (P=0.007) or TNM stages (P=0.022). Five-year recurrence-free survival time (P<0.001) and overall survival time (P<0.001) in patients with high CTEN expression were signiifcantly less than those in patients with low CTEN expression. Multivariate Cox regression analysis revealed that the CTEN expression was an independent prognostic marker for HCC (all P<0.05). Conclusion: CTEN protein may play a role in the genesis and development of HCC, and it can function as a prognostic marker.

Objective To investigate the risk factors and predictive model for the occurrence of post-sustained virologic response (SVR)hepatocellular carcinoma in chronic hepatitis C (CHC)patients. Methods A total of 203 CHC patients hospitalized at the First Affiliated Hospital of Zhengzhou University from January 2006 to December 2014 who received antiviral therapy and achieved SVR were collected,including 11 post-SVR HCC cases.Risk factors for post-SVR HCC were estimated by Cox′s proportional hazards regression model.Cutoff value predicting risk of post-SVR HCC was determined by receiver operating characteristic curve.Results In Cox′s model,the risk of post-SVR HCC increased by 9.4-fold in patients with initial diagnosis as compensated cirrhosis compared to those with initial diagnosis as CHC.Increase in post-SVR albumin by per 1 g/L was associated with reduced risk by 20% for the occurrence of post-SVR HCC.Cut-off value of post-SVR albumin for the prediction of HCC was determined as ≤ 36.0 g/L with an area under the curve (AUC)of 0.809.A predictive model for post-SVR HCC was created based on initial diagnosis as compensated cirrhosis and post-SVR albumin ≤36.0 g/L with an AUC of 0.871 .The sensitivity,specificity and negative predictive value of the model were 0. 818,0.896 and 0.989,respectively.Conclusions Initial diagnosis as compensated cirrhosis combines with post-SVR albumin ≤36.0 g/L are risk factors for post-SVR HCC with ideal prediction value for the occurrence of post-SVR HCC in CHC patients.

Objective To apply special staining techniques in pathological diagnosis of fungal infections in HIV/AIDS patients.Methods Pathological data of 20 HIV/AIDS patients complicated with fungi infections in Shanghai Public Health Clinical Center during February 2010 and November 2013 were retrospectively analyzed.Tissue specimens were stained with hematoxylin and eosin (HE),Periodic acidSchiff (PAS) and methenamine silver nitrate (MSN),and the sections were observed under optical microscope.Results Among 20 HIV/AIDS patients complicated with fungi infections,2 were infected with pulmonary cryptococcosis; 3 were penicillium marneffei infections in skin,lung and abdominal mesenteric lymph nodes; 5 were histoplasma capsulatum infections in epiglottis,neck lymph nodes,oral cavity,abdominal cavity and skin; 4 were aspergillus infections in maxillary sinus,lung and vocal cords,and 3 of them were combined with tuberculous lesions; 6 were candida albicans infections in liver,pharynx,esophagus and stomach.In tissues stained with HE the infiltration of inflammatory cells,granuloma formation and coagulative necrosis were observed,and the shape of fungi needed careful observation to avoid missed diagnosis and misdiagnosis.In tissues stained with PAS,fungal spores and pseudohypha were presented in bright amaranth,and cell nucleus was in purple-blue.In tissues stained with MSN,fungal spores and pseudohypha were identified clearly in brown-black.Conclusion HE plus PAS and MSN staining will help pathological diagnosis of fungi infection.

Objective To understand the changes of cardiac systolic and diastolic function in human immunodeficiency virus (HIV)-infected patients without evidence of cardiac disease in China.Methods Forty-two HIV-infected patients who were followed up in the Department of Infectious Diseases at Peking Union Medical College Hospital without cardiac involvement were recruited.All the HIV-infected patients had received highly active antiroviral therapy (HAART) for more than 12 months with viral suppression.And 30 age and sex matched healthy subjects without cardiac disease manifestations were enrolled as controls.Every group members underwent transthoracic echocardiography evaluation.The indexes of cardiac systolic and diastolic function between HIV-infected patients and healthy controls were compared.Results Diastolic abnormality occurred in 20 cases in HIV-infected group and 6 cases in control group, with statistically significant difference (χ2=5.79, P=0.007).The E wave deceleration time (EDT) in HIV-infected patients were significantly decreased than healthy controls ([161.87±21.64] ms vs.[190.34±37.22], t=-3.20, P=0.002).There were no significant differences of E/A ratio ([1.16±0.35] vs.[1.19±0.26]), E/Ea ratio ([5.43±1.99] vs.[5.78±0.91]), isovolumic relaxation time (IVRT), ([93.18±20.34] ms vs.[93.57±18.55]ms), Ea ([10.18±2.80] cm/s vs.[11.45±2.75] cm/s) between HIV-infected patients and controls (t=1.13,1.53,0.67 and 0.29, respectively, all P>0.05).Among cardiac systolic function markers, left ventricular ejection fractions in HIV-infected patients and control group were (66.7±6.4)% and (68.7±4.2)%, respectively.And left ventricular shortening rates were (37.08±4.79)% and (38.17±3.96)%, respectively.Both showed no significant difference between the two groups (t=-1.51 and-1.00, respectively, both P>0.05).Conclusions Compared with control group, subclinical cardiac diastolic dysfunction is more frequently observed in HIV-infected patients.However, there are no significant differences of cardiac systolic function markers between HIV-infected patients and controls.

Objective To analyze the abnormal detectable rates of different kinds of blood test indexes before and after treatment in the patients with snake bite and to probe into the change condition of these indexes after different snake bite .Methods The inpa‐tients with clearly diagnosed what kind of snake bite in the past two years were selected as the research subjects .The multiple blood test indexes were determined before and after treatment in these patients .The abnormal detectable rates of these indexes were ana‐lyzed and compared among various snake bite patients .Results The different kinds of snake bite all could cause the different de‐grees of changes in some detection indexes among partial patients .Specially ,the detectable rates of WBC ,PT ,APTT ,TT ,D‐D ,CK , CK‐MB ,LDH ,Urea ,Cr and Cys‐C increase and PLT and Fbg decrease caused by viper bite were apparently higher than those caused by other kinds of snake bites (P<0 .05) .The detectable rates of CO2 increase and K+ decrease in the coral snake bite were apparently higher than those in trimeresurus gramineus ,cobra and viper bite (P<0 .05) .Conclusion Because the snake species and toxicities are different in the various snake bites ,so the caused changes and the abnormal detectable rates of blood test indexes also are different .