ObjectiveTo investigate the expression of midkine(MK) and its relation with angiogenesis, biological features and prognosis of pancreatic carcinoma(PC).MethodsMK expression and microvessel density(MVD) were determined in 52 cases of human PC with immunohistochemistry and results were compared with pathology.ResultsMean MVD of PC was 64?18 and positive expression of MK was detected in 38 cases (73%). The positive rate of MK was significantly lower in cases of without metastasis and at early clinical stage (stageⅠ～Ⅱ) than that with metastasis and at stage Ⅲ～Ⅳ. MVD was significantly higher in MK-positive PC than in MK-negative PC (P

Objective To investigate the biological effect of transforming growth factor-?1(TGF-?1) on inducing myofibroblast formation in hypoxic pulmonary vascular remodeling.Methods Forty male Wistar rats were exposed to hypoxia for 0,3,7,14 or 21 days.Mean pulmonary arterial pressure(mPAP),vessel morphometry,right ventricle hypertrophy index(RVHI) were measured.Immunocytochemistry was used to measure the expression of ?-Smooth-muscle actin(?-SMA) and TGF-?1 in pulmonary artery walls and in situ hybridization was used to measure the expression of TGF-?1 mRNA in pulmonary artery walls.Ultrastructure alveolar wall vessels were observed by electron microscopy.Human embryonic lung fibroblasts(KMB17) phenotype after induction of hypoxiaand TGF-?1 were recorded through cell culture.Results(1) mPAP increased significantly after 7-day of hypoxia(P

Growth hormone (GHRH) and pitutary adenylate cyclase activating peptide (PACAP) are the members of the PACAP/Glucagon superfamily,who are related in both sequence and function.Their stimulation of GH secretion and animal growth is concerned.A series of expression plasmid,pIRES1-GHRH-PACAP (P-G-P),plRESI-GHRH (P-G) and plRESI-PACAP(P-P),were constructed,extracted and purified,then transfected into CHO cell line with Lipofectamine.The expression was examined by RT-PCR,dot-ELISA and Western blotting.The biological activity of expression products was detected in rats.At 8 h after injection of transfection supematant,serum IGF-I concentrations in P-G-P group were significantly higher than that in other groups(P ＜ 0.05).PLGA encapsulating plasmid microspheres were prepared and injected intramuscularly into rabbit legs.Growth behavior and IGF-1 level were measured at day 0,15,30 and 45 after injection.Greater body weights gain and higher serum 1GF- [ levels were observed in three plasmid microsphere injection groups,compared with control group.At day 30,the body weight gain in P-G-P group was greater than saline group (81%,P＜ 0.01),P-G mierosphere group (15%,P＜ 0.05) and P-P microsphere group (7%,P＞ 0.05),serum IGF-I concentration in P-G-P microsphere group showed a 16.68% increase to P-G microsphere (P ＞ 0.05),a 17.14% increase to P-P microsphere(P ＞ 0.05) and a 50.46% increase to control (P ＜ 0.05).These results suggest that co-expression of GHRH and PACAP in one expression plasmid might exert an additive stimulation of GH secretion and growth when delivered into rabbit skeletal muscle with PLGA mierosphere.The results may provide a new approach to regulate animal growth.

Objective To investigate the effect of liver depression (Liver Qi Stagnation) on Th17,Treg,IL-17,IL-10 and airway inflammation in asthmatic rats,and to clarify the immune mechanism of asthma with liver depression.Methods Established the combined with disease and syndrome model of asthma with liver depression.Collected the bronchoalveolar lavage fluid (BALF) to count the total and differential cell.Lung tissue was observed in microscope; the proportion of Th17 cells and Treg cells of CD4 +T cells in peripheral blood was measured by flow cytometry; the levels of IL-17 and IL-10 were determined by ELISA.Results The total number of inflammatory cells[(96.86±4.43)× 107/L,(88.22±3.22)× 107/L],the proportion of eosinophils [(27.58 ±4.65) ％,(22.67±2.43) ％],Th17 cells[(6.86±0.98) ％,(6.01 ±0.77) ％] and IL-17 level [(48.88± 8.06)pg/ml,(43.24± 6.32) pg/ml] of asthma in liver depression group and asthma group were significantly higher than the control group [(30.58 ± 2.49) × 107/L,(0.78 ± 0.12) ％,(2.80± 0.82) ％,(24.11 ±3.40)pg/ml]; Treg cells [(3.09±0.55) ％,(3.96±0.66) ％] and IL-10 level [(19.79±2.80) pg/ml,(20.29±3.12) pg/ml] were significantly lower than the control group [(8.02± 1.26) ％,(30.79 ± 4.01) pg/ml].The total number of inflammatory cells (96.86 ±4.43) × 107/L,the proportion of eosinophils (27.58±4.65) ％ and Th17 cells(6.86±0.98) ％ and IL-17 level (48.88±8.06)pg/mL of asthma in liver depression group were significantly higher than the asthma group (88.22 ± 3.22) × 107/L,(22.67 ± 2.43) ％,(6.01 ± 0.77) ％,(43.24 ± 6.32) pg/ml;the proportion of Treg cells (3.09 ±0.55)％ was significantly lower than the asthma group (3.96± 0.66)％; and the lung histopathology symptoms was more severe than asthma group.Conclusion Liver Qi Stagnation can promote the inflammation of asthma,the imbalance of Th17/Treg and IL-17 level to aggravate the asthma.Liver depression is one of the major internal factors in recurrent episodes of asthma.

Objective To investigate the relationship among the concentration of plasma interleukin (IL)-10 and plasma IL-13 and the clinical therapeutic effect in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods Thirty-six AECOPD inpatients were enrolled in this study.Blood samples of the subjects were collected as soon as hospitalization, and plasma IL-10 and IL-13concentrations were analyzed by enzyme-linked immunosorbent assay.The clinical manifestations of subjects were quantified by a special designed score standard, and were evaluated at the time points of hospitalization, 48 hours treatment and 96 hours treatment.The therapeutic effect was evaluated by clinical manifestations score combined with pulmonary ventilation functional parameter.Eventually, the correlation among the concentration of IL-10 concentration and IL-13 and the clinical therapeutic effect were analyzed.Results The correlation coefficients between clinical manifestations score decrease and pulmonary ventilation function improvement with plasma IL-10 concentration after 48 hours treatment were 0.85 and 0.48 respectively,then, 0.64 and 0.52 after 96 hours treatment.The correlation coefficients between clinical manifestations score decrease and pulmonary ventilation function improvement with plasma IL-13 concentration after 48hours treatment were -0.41 and -0.34, after 96 hours treatment , correlation coefficients between clinical manifestations score decrease with plasma IL-13 concentration was -0.36.All of the above correlation coefficients were statistically significant.Conclusion Plasma IL-10 concentration was positively, whereas, IL-13 concentration was negatively correlated with the therapeutic effect in AECOPD patients.

Objective To establish a multiple PCR method that can be used to spontaneously detect five kinds ofpathogens such as NG,MH,MG,CT and UU.Method With the fluorescence-quantitative PCR technique in conjunction with another detection technique as the golden standard,evaluation was conducted on the sensitivity,specificity,accuracy and repeatability on the detection of 5 kinds of STD pathogens using single-tube multiplex PCR.Result The sensitivity,specificity and match rate of the method ale 10-9fg/μl,100%,97.8%respectively,and the repeatability of 5 continuous days of 20 clinical specimens is good.Conclusion Single-tube multiplex PCR technique provides a new method to detect 5 kinds of STD pathogens.

Objective To observe the influence of suspension concentrate of niclosamide on the enzyme activity of Oncomelania hupensis in order to explore its molluscicidal mechanism. Methods Oncomelania hupensis snails were collected in the habitates of river marshland in Dantu District, Zhenjiang City, Jiangsu Provence and were divided into 2 groups. The snails of the treated group were sprinkled with 25% suspension concentrate of niclosamide. The snails of the control group were sprinkled with distilled water. The soft body tissue of the snail was separated and the sections of snail tissue were made in the Cryostat Microtome. The stain of enzyme-histochemistry showed CCO, LDH, SDH, CHE and NOS had been done, and then the staining block was made by routine method. The staining reaction in the snail tissue and the average gray density were observed with the image analysis system of biomicroscope. Results The enzyme activity of CCO, LDH, SDH, CHE and NOS located in the mouth, muscle fiber, tegumentary membrane, ganglia, liver and pharyngeal cavity of Oncomelania hupensis snails. The enzyme activities of CCO, LDH, SDH, CHE and NOS in the treated group were significantly lower than that in the control group. Conclusions Niclosamide can affect the transmitting of neurohypophysis and obstruct the energy and result in the disorder of the physiological functions in Oncomelania hupensis. It is one of the reasons of Oncomelania hupensis death.

Objective To explore the Schistosoma japonicum cercariae collected method and the quantity of cercariae obtained from a great quantity artificial infected snails. Methods In laboratory condition, Oncomelania snails were infected with schistosome miracidia. Sixty days post-infection all snails were divided into 7 shares. Cercariae shed from 1 share snails every day and the number of all shedding days were 40. Cercariae shed from snails were collected with low-velocity centrifuge and the cercariae sediment were weighted. Results One thousand and nine hundred g snails bred for 120 days post-infection, the infection and survival rates were 36. 00% and 51. 58%. Cercariae col-lected were 10. 5 g from 40 days collection. Cercariae quantity of shedding from 1 000 positive snails per day was 0. 257 4 g.

Objective To observe the impact of water temperature on shedding and infectivity of the cercariea of Schistosoma japonicum under laboratory condition. Methods Infected snails were exposed to 1, 5, 10, 15, 20, 25, 30, 35℃ and 40℃ water for 4 hours for shedding of cercariae respectively, the shedding rates and total numbers of cercariae shed were investigated. The mice were infected with cercariae in 1, 10, 20, 30℃ and 40℃ water for 30 minutes respectively, the infection rates and mean worm burden of mice were investigated. Results The cercariae were shed from infected snails with Jiangsu or Yunnan isolate of [WT5”BX]S. japonicum in water ranged from 1℃ to 40℃, but the fittest water temperatures range for shedding of cercariae of Yunnan isolate of S.japonicum[WT5”BZ] is from 20℃ to 35℃, while fittest range for Jiangsu isolate is from 15℃ to 35℃. All of the infection rates of mice infected with cercariae of Jiangsu isolate of S.japonicum in the water ranged from 1℃ to 40℃ were 100.0%. Infection rates of mice infected with cercariae of Yunnan isolate of S. japonicum in the water ranged from 1℃ to 40℃ were more than 83.3%. Conclusions When infected snails are exposed to the water with temperature ranged from 1℃ to 40℃ it is possible for cercariae to shed and infect the final hosts . It is suggested that there is a possibility of infection with S.japonicum when contacting the water near marshlands with infected snails in winter.

Objective To find out the valuable early diagnostic antigen of Schistosoma japonicum. Methods The sera of rabbits were collected at different time after the rabbits were infected with cercariae of Schitosoma japonicum. The fractions of the soluble cercaria antigen (SCA), soluble adult worm antigen (AWA) and soluble egg antigen (SEA) were separated by SDS-PAGE and recognized by Western blotting with rabbits' sera of different time of post-infection. Results In Western blotting, the bands of 94, 48, 41, 40 kDa and 38 kDa of SCA appeared the earliest and were recognized by the rabbits sera of 2-week post-infection, the bands of 71 kDa and 23 kDa of SCA reacted with the rabbits sera of 3-week post-infection strongly. The bands of 71 kDa and 58 kDa of AWA appeared the earliest and were recognized by rabbits sera of 3-week post-infection. The bands of SEA reacted earliestly to the rabbits sera of 4-week post-infection were 270, 151, 73, 69, 50 kDa and 24 kDa. Conclusion The fraction antigens of 94, 71, 48, 41, 40, 38 kDa and 23 kDa of SCA, the fraction antigens of 71 kDa and 58 kDa of AWA and the fraction antigens of 270, 151, 73, 69, 50 kDa and 24 kDa of SEA could be recognized by sera of acute infected rabbits and might have potential early immuno-diagnosis value for schistosomiasis.