Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of heme oxygenase-1 ( HO-1 ) in THP-1 cells and its possible mechanism .Methods Human monocyte cells THP-1 were cultured in vitro and then were incubated with various concentrations (0, 0.01, 0.1, 1.0 or 5.0 ng/ml) of MALP-2 for 16 h, or were stimulated by 5.0 ng/ml MALP-2 for different length of time (0 h, 4 h, 8 h, 12 h, 16 h or 24 h).The expression of HO-1 at mRNA and protein levels were detected by real-time PCR analysis and Western blot assay .The enzyme activity of HO-1 was detected by colorimetric analysis.THP-1 cells were pre-incubated with 30 μmol/L of SB203580, PD98059 and SP600125 for 30 min and then were cultured with 5.0 ng/ml MALP-2 for 16 h to investigate the role of mito-gen-activated protein kinases (MAPKs) signaling pathway in HO-1 production.After incubating THP-1 cells with 5.0 ng/ml MALP-2 for different periods of time, NF-E2-related factor 2 (Nrf2) protein was detected by Western blot assay to study the effects of Nrf2 pathway on MALP-2-induced HO-1 expression.Nrf2 and HO-1 proteins were measured by Western blot assay after transfecting THP-1 cells (1×106/well) with Nrf2 siRNA at a final concentration of 100 nmol/L.Results MALP-2 enhanced the expression of HO-1 at mRNA and protein levels as well as the enzyme activity of HO-1 in THP-1 cells in a concentration-dependent manner.The expression of HO-1 protein induced by MALP-2 was significantly inhibited by 30 μmol/L MAPKs specific inhibitors ( SB203580 , PD98059 and SP600125 ) .MALP-2 induced Nrf2 translocation at a concentration of 5.0 ng/ml.The expression of Nrf2 and HO-1 proteins were significantly decreased in Nrf 2 siRNA-transfected THP-1 cells.Conclusion MAPKs and Nrf2 signaling pathways were involved in the MALP-2 induced HO-1 expression .

Objective:To observe the molecular mechanism involved in expression of hemeoxygenase -1 (HO-1) induced by a macrophage-activating lipopeptide-2 (MALP-2).Methods:THP-1 cells were cultured in vitro and stimulated by MALP-2 for 12 h, expression of HO-1 was detected by Western blot .TLR2 and TLR6 neutralizing antibodies incubation , dominant negative plasmids transfection were used to assess the functional of TLR 2,6 in mediating HO-1 expression.Phosphorylation of c-Src and Akt were detec-ted by Western blot, and c-Src siRNA and PI3K inhibitor LY294002 were used to investigate the role of c-Src and PI3K in HO-1 ex-pression.Results:MALP-2 induced c-Src phosphorylation , and TLR2 and TLR6 neutralizing antibodies , or their dominant negatively plasmids could abrogate this effect .In addition, siRNA of c-Src could decrease the phosphorylation level of Akt , and the PI3K inhibi-tor could inhibit HO-1 expression.Conclusion: MALP-2 can induce THP-1 cells expression of HO-1 through TLR2,6/c-Src/PI3K pathways .

Objective To measure total hemoglobin concentration (THC) of breast lesion using US-guided diffused optical tomography(DOT) and to investigate the THC optimal threshold value in different size breast lesions.Methods DOT was performed on 500 breast lesions and surgical pathology was as the gold standard.The optimal diagnostic threshold and the efficacy were figured out.Results There were 265 benign and 235 malignant lesions.In malignant lesions,THC of ≥2 cm lesion group was higher than that of ＜2 cm lesion group(P =0.000).In benign lesions,there was no statistical difference between ≥2 cm group and ＜2 cm group (P =0.13).As for ＜2 cm breast lesions,when a THC threshold value of 146.9 μmol/L and 102.2 μmol/L were used,the sensitivity,specificity,positive predictive value,negative predictive value and accuracy were 74.2％,70.0％,71.7％,62.9％,79.9％ and 86.7％,44.4％,61.6％,51.6％,83％,respectively.As for ≥2 cm breast lesions,when a THC threshold value of 210.4 μmol/L were used,the sensitivity,specificity,positive predictive value,negative predictive value and accuracy were 74.0％,86.7％,79.1％,89.2％,69.2％.Conclusions THC of breast cancer increased with the increasing size of lesions.The different diagnostic threshold value should be used according to different size lesions so as to enhance sensitivity,specificity and accuracy.

Objective To formulate a five-level pediatric emergency triage standard and evaluate the efficacy and reliability of it in determining severity of emergency pediatric patients.Methods According to the assessment methods in Pediatric Advanced Life Support recommended by American Heart Association and American Academy of Pediatrics,we formulated a five-level pediatric emergency triage standard based on the situation of our hospital and analyzed the data one year before and after the application of it.Results Before and after the application of the triage standard,the average satisfaction rate of emergency patients were (81.28 ± 3.97) ％ and (94.13 ± 4.62) ％,and there was significant difference (P ＜ 0.01) ; the proportion of whom became worse during waiting time were 1.83％ (628/34275) and 0.04％ (16/36 187),and there was significant difference (P ＜0.01) ;the average waiting time of emergency admission patients were (12.71 ± 2.32) min and (3.34 ± 1.95) min,and there was significant difference (P ＜0.01) ;the misjudgment rate of severity were 3.78％ (1 296/34 275) and 0.57％ (205/36 187),and there was significant difference (P ＜ 0.05).Conclusion The five-level triage standard is objective,easy to master,and suitable for pediatric triage.It can quickly sort out critical cases from emergency pediatric patients,which can improve the effectiveness of emergency service,make use of medical resources rationally and somewhat solve the problem of overcrowding.

Objective To evaluate the inter-observer variability of static breast sonogram final assessment among observers with different breast imaging experience, using the first edition of the Breast Imaging Reporting and Data System(BI-RADS) for ultrasound. Methods Thirty patients with 30 breast lesions were included who underwent beast lesions resection operation. A pathological diagnosis was available for all 30 lesions:16 (53%) malignant and 14 (47%) benign. Twelve radiologists independently reviewed two sonograms of each lesion, and assigned a final BI-RADS assessment category. Inter-observer variability was measured using kappa statistic. Positive predictive value(PPV) and negative predictive value (NPV) for final assessment were also calculated. Results As for the experienced observers,kappa values of categories 3,4 and 5 were 0.72,0.28 and 0.60,NPV of category 3 was 93% ,PPV of category 5 was 97% ,all of which decreased as the breast imaging experience reduced. PPVs of categories 4a,4b and 4c were 56 % ,88% and 69%, respectively. Conclusions Using BI-RADS final assessment, radiologists with sufficient breast imaging experience can provide accurate and consistent assessment for breast ultrasonography,but the agreement of diagnosis decreased as the breast imaging experience reduced. The clinical feasibility of 4a,4b and 4c subcategories is uncertain.

OBJECTIVE To established a viable and convenient method for human nasoseptal chondrocyte culture in vitro and study the biological characters of the chondrocyte. METHODS The chondrocytes were obtained by incomplete enzyme digestion of cartilage and were seeded with small piece of cartilage in 25 ml culture flasks coated with polylysine previously. This study determines the biologic synthesis of the chondrocytes by toluidine blue and type Ⅱcollagen immunohistochemistry reaction. The proteoglycan synthesis efficiencies of human nasoseptal chondrocytes cultured in vitro were measured by using 35S-Na2SO4 incorporation. The modified method and the conventional method in proteoglycan synthesis were also compared. RESULTS The results suggested that human nasoseptal chondrocytes cultured by modified method still show the normal structure and morphology, and maintain the function of metabolism and self duplication. With the modified method, we can get a higher cell survival rate and more cells. CONCLUSION The modified method is a viable and convenient method.

Objective To study the reasons of natural death of Oncomelania hupensis snails in marshland by comparing the differences of environmental factors between snail marshland and non-snail marshland in East Dongting Lake schistosomiasis en-demic areas. Methods Two adjoining marshlands were selected,one was a snail marshland and another was a non-snail marsh-land. The measuring points were set by grid of“20m × 20m”. The soil moisture,elevation,soil pH and temperature of the points were measured,and the snails of the points were surveyed. Results The elevation,soil pH and temperature of the snail natural death marshland were all lower than those of the snail marshland(P<0.01). The soil moisture,elevation,soil pH and temperature of different snail densities in the snail marshland were different(P<0.05). The soil moisture of the non-snail group was higher than that of other four groups(P<0.01)and the soil moisture of the group with 0-11 snails per frame was lower than that of other four groups(P<0.01). The elevation of the group with 0-11 snails per frame was lower than that of the group with 24-39 snails per frame and the group with more than 39 snails per frame(P<0.05). The soil pH of the group without snails was higher than that of other four groups(P<0.01)and the soil temperature of the group with 0-11 snails per frame was lower than that of the group with 24-39 snails per frame. The differences of soil moisture and elevation of different death snail densities were not significant, while the differences of soil pH and temperature were significant. The soil pH of the non-snail died group was higher than that of the 2-died,3-died and more than 3-died groups(P<0.05). The soil temperature of the non-snail died group was higher than that of other four groups(P<0.05). Conclusion The elevation,soil pH and soil temperature within the range of the research may be the factors affecting the snail natural death and the soil moisture,elevation,soil pH and soil temperature may be the factors affect-ing the distribution of snails.

Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of hemoxygenase-1 ( HO-1 ) in THP-1 cells and to further elucidate its possible regulatory mechanism for a better understanding of protective response upon mycoplasma infection .Methods THP-1 cells were cultured in vitro and stimulated by MALP-2 at different concentrations for 12 h.THP-1 cells were incubated with TLR 2 or TLR6 neutralizing antibodies , or transfected with their dominant negative plasmids to evaluate the effects of TLR 2 and TLR6 on HO-1 expression .Phosphorylation of Akt was detected by Western blot.PI3K inhibitor LY294002 was used to investigate the role of PI3K in HO-1 expression.Im-munofluorescence and electrophoretic mobility shift assay ( EMSA ) were performed to observe the nuclear translocation and DNA-binding activity of nuclear factor Nrf 2.Small interfering RNA ( siRNA) was used to silence the genes encoding Nrf2 and HO-1.Cobalt protoporphyrin (CoPP), an inducer of HO-1, was used to treat THP-1 cells.The expression of HO-1 was detected by Western blot .The secretion of TNF-αand IL-1βby THP-1 cells were measured by ELISA .Results MALP-2 induced the expression of HO-1 in THP-1 cells.However, the expression of HO-1 was inhibited by TLR2 and TLR6 neutralizing antibodies and expres-sion of their dominant negative plasmids .Moreover, PI3K pathway was activated by MALP-2, and with the use of PI3K inhibitor, the expression of HO-1 was decreased.The translocation of Nrf2 to the nucleus and itsDNA-binding activity were enhanced by MALP-2, but were inhibited by the treatment of PI3K inhibitor.Theexpression of HO-1 was significantly down-regulated upon the interference of Nrf2 gene expression withsiRNA.Silenced expression of HO-1 increased the level of TNF-αand IL-1β, while CoPP treatment decreasedthe secretion of MALP-2-induced cytokines.Conclusion MALP-2 might induce the expression ofHO-1 in THP-1 cells through TLR2,6/PI3K/Nrf2 pathways.The expression of HO-1 could negatively regulatethe hyper-secretion of cytokines.

Objective To understand the distribution of univalvia molluscs in Eastern Dongting lake area where the Oncome-lania hupensis is extinct,so as to explore the causes of extinction and to provide the evidence for formulating schistosomiasis con-trol strategy. Methods The univalvia molluscs of the Qianliang Lake district,Jianxin District and Junshan Park were investigat-ed in August of 2013. All the collected snails were classified and identified. The data were analyzed by using the method of Shapiro-Wilk normality test and non-parametric test. The ecological biodiversity differences from each district were compared by the biodi-versity indexes. Results The univalvia molluscs collected were from 6 species in 3 families in Gastropoda class,namely Oncome-lania hupensis Gredler in Pomatiopsidae family,Parafossarulus striatulus,Alocinma longicornis,Parafossarulus sinensis,Bithynia fuchsisana in Bithyniidae family,and one genus provisionally named Radix in Lymaneidae family. In Junshan Park ,a total of 4 553 snails were collected,among which 1 264 were Oncomelania snails. In Jianxin District,336 univalvia molluscs were collect-ed,and no Oncomelania snails were found. In Qianliang Lake district,there were only 7 Alocinma longicornis snails,2 Parafossa-rulus sinensis snails and 1 Parafossarulus striatulus snail collected. There were significant differences among the distributions of the samples from the three districts(χ2=166.225,P<0.01).The Simpson’s diversity indexes in Qianliang Lake area,Jianxin Dis- trict and Junshan Park were 0.402 8,0.718 6 and 0.678 8,respectively,and the Shannon-Wiener indexes of the 3 areas were 0.721 5,1.431 3 and 1.199 9,respectively. Conclusions With the extinction of Oncomelania snails,the species and quanti-ties of other snails become rare in Qianliang Lake area. Whether their causes are relevant is worth further studying.

Objective To study the reasons of natural death of Oncomelania hupensis snails by comparing the differences of the indicator days covered with water DCW in snail marshland and non?snail marshland around the build of Three Gorges Dam in Eastern Dongting Lake areas. Methods Two marshlands were selected one was a non?snail marshland Qianliang Lake and another was a snail marshland Junshan Park . The measuring points were set through the mechanical sampling. The snails and elevation of the points were surveyed and the data of the water levels from the hydrological station were collected and then DCWs were calculated. Results From 1995 to 2013 DCWs of the marshland of natural death of snails were all more than that of the snail marshland P<0.01 . In Qianliang Lake marshland the difference between DCW before natural death and DCW from natural death until the dam was not significant P=0.23 while DCWs of the two stages both were more than that after the dam P1=0.045 P2=0.002 . Before the build of the dam DCW of the Qianliang Lake marshland of natural death of snails was more than that after the build of the dam P=0.013 and there was the same situation in Junshan Park marshland P=0.005 . The relationship between snail density and DCW was not significant in Junshan Park marshland rs=0.008 P=0.914 and the reference range of DCW of all the measuring points was 76-251 days. Conclusion In the eastern Dongting Lake district the build of Three Gorges Dam and DCW may be not the direct factors affecting the natural death of snails and the latter may change the distribution of snails.