Objective To prepare recombinant mda-7/IL-24 adenovirus to study its function on the proliferation of small cell lung cancer NCI-H446 cells. Methods According to the manufacturer's instructions of AdEasy vector system, mda-7/IL-24 cDNA was subcloned into the adenoviral shuttle vector pAdTrack-CMV. The efficient recombination of adenoviral backbone vector pAdEasy-1 and pAdTrack-CMV-mda-7/IL-24 was achieved in bacteria E. coli BJ 5183. The recombinant adenoviral vector pAd-mda-7/IL-24 linearized by Pac Ⅰ was transfected into HEK293 cells with lipofectamine 2000. To generate higher titer viral stocks, the amplification of recombinant adenovirus was accomplished in packing cells. Viral titers were measured by tissue culture infectious dose 50 (TCID_ 50 ) method. Ad.mda-7/IL-24 was identified by PCR. The expression of pAd-mda-7/IL-24 in NCI-H446 cells was detected by Western-blot analysis. The function of Ad.mda-7/IL-24 on the proliferation of NCI-H446 cells was assayed by MTT after cells were infected by 50 pfu/ml adenovirus. Results The recombinant adenoviral shutter vector pAdTrack-CMV-mda-7/IL-24 and recombinant adenoviral vector pAd-mda-7/IL-24 were constructed successfully as identified by sequence analysis. PCR assay showed that adenovirus Ad.mda-7/IL-24 contained mda-7/IL-24 cDNA. After amplification in packing cell HEK293, the titer of virus was 2?10~ 10 pfu/ml measured by TCID_ 50 assay. Western-blot results identified that MDA-7/IL-24 could be expressed in NCI-H446 cells. After infected by 50 pfu/ml adenovirus, the proliferation of NCI-H446 cells was inhibited by 21.37% with MTT method. Conclusion Ad.mda-7/IL-24 can inhibit the growth of NCI-H446 cell obviously. This result lays foundation to study its function mechanism and to apply it in gene therapy of the small cell lung cancer.

Objective To explore the role of plasminogen activator inhibitor-1(PAI-1)in development of progressive fibrosis via the inhibition of extracellular matrix degradation,and to reveal the contributive role of PAI-1 in muscular dystrophy(MD).Methods Expression and cellular localization of PAI-1 protein were examined in frozen muscle specimens obtained via biopsy from 5 patients with duchenne muscular dystrophy(DMD),3 patients with becker muscular dystrophy(BMD),9 patients with congenital muscular dystrophy(CMD) and 4 cases with normal muscle by immunohistochemistry,double immunofluorescence and Western-blot analysis.Results PAI-1 was positive only in vascular endothelial cells of normal muscle.Both immunohistochemistry and Western-blot analysis showed that PAI-1 expression distinctly increased in most dystrophic muscles of MD than that in normal muscles.Double immunolabeling revealed that PAI-1 strongly expressed in cytoplasm and nuclei of regenerating muscle fibers,macrophages,macrophage infiltrating necrotic fibers.Some activated fibroblasts in endomysium and perimysium of DMD and CMD muscles were positive for PAI-1.Conclusions The functional consequence of overexpression of PAI-1 in dystrophic muscles is unknown but the elevated local expression of PAI-1 in diseased muscles of MD and their distinct distribution pattern provide evidence that PAI-1 participate in pathogenesis of MD.

OBJECTIVETo examine serum concentration of interleukin-18 (IL-18) and IL-18 mRNA expression in peripheral blood mononuclear cells (PBMCs) in children with primary nephrotic syndrome (PNS) and explore the possible role of IL-18 in steroid-resistant nephrotic syndrome (SRNS).

METHODSSixty-six children with newly diagnosed PNS, including 39 cases of steroid sensitive nephrotic syndrome (SSNS) and 27 cases of SRNS, were enrolled. Forty healthy children were used as a normal control group. Blood samples were collected before and 8 weeks after glucocorticoid treatment. Serum concentration of IL-18 was measured using ELISA. IL-18 mRNA expression in PBMCs was detected by the RT-PCR method. The amount of 24-hr urine protein was measured by the biuret method. Serum contents of total cholesterol (T-Ch), triglyceride (TG), low density lipoprotein (LDL), total protein (TP), and albumin (Alb) were measured by the automatic biochemistry analyzer.

RESULTSSerum concentration of IL-18 and IL-18 mRNA expression in PBMCs in the SSNS and the SRNS groups were significantly higher than those in the normal control group before treatment (P< 0.05). The SRNS group had increased serum protein concentration of IL-18 and IL-18 mRNA expression in PBMCs compared with the SSNS group before treatment (P< 0.05). Serum LDL content in the SRNS group was also significantly higher than that in the SSNS group before treatment (P< 0.05). After treatment, serum concentration of IL-18 and IL-18 mRNA expression in PBMCs in the SRNS group were significantly higher than those in the SSNS and the normal control groups (P< 0.05). Serum concentration of IL-18 and IL-18 mRNA expression in PBMCs in the SSNS group were significantly reduced after treatment, but the alterations of IL-18 were not observed in the SRNS group after treatment.

CONCLUSIONSSRNS was associated with increased serum IL-18 concentration and IL-18 mRNA expression in PBMCs. Over-production of IL-18 may play a role in the development of SRNS.

Adolescent ; Adrenal Cortex Hormones ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance ; Female ; Humans ; Interleukin-18 ; blood ; genetics ; physiology ; Leukocytes, Mononuclear ; metabolism ; Lipoproteins, LDL ; blood ; Male ; Nephrotic Syndrome ; blood ; drug therapy ; RNA, Messenger ; blood

Objective To evaluate the eff ec t of snail control with niclosamide ethanolamine salt dustable powder (DP) in th e marshland of the Yangtze River ahead of the schedule in spring, and to underst and the impact of the low temperature to molluscicidal effect of the dusting. Methods The snail control test with niclosamide ethanolamine salt dustable powder was carried out under different te mperatures in laboratory. Then the snail control was undertaken in the marshland of the Yangtze River ahead of the schedule in spring, and the molluscicidal ef fect was observed and the temperature changes were also considered. Results When the temperature was 15 ℃ and 25 ℃ respectively, there was no significant difference on the molluscicidal effec t of niclosamide ethanolamine salt DP in laboratory. The average atmosphere temp erature in each day was 8.9-16.3 ℃ from the last ten-day of March to the fir st ten-day of April in Nanjing regions. The corrected mortality of snails and the r educed rates of density of snails were 95.38% and 99.97% respectively on the s ur face of the soil in the marshland field. The corrected mortality and the reduced rates of density of snails were 79.30% and 70.59% respectively in the surface l ayer of the soil. Conclusion The impact of air t emperature is little to the molluscicidal effect of niclosamide ethanolamine sal t DP when the average atmosphere temperature in each day was 10-15 ℃ in the ma r shland of the Yangtze River. If the snail control is undertaken ahead of the sch edule in spring, the reproduction of snails will be suppressed and the infection of Schistosma japonicum will be controlled. [

Objective To determine the de ve lopmental zero and effective accumulated temperature of Oncomela nia eggs under con stant temperature. Methods The eggs were cultur ed under different constant temperature, and their development duration and effe ctive accumulated temperature were studied. The zero accumulated temperature (℃ ) was obtained by using an estimation mode, and the effective accumulated temp erature was obtained by effective accumulated mode of K=N(T-C). [WT5”H Z] Results The average development duration was (27.29?17 .2 9) d under 15-30 ℃, the development zero temperature was 11.79 ℃ and 38.22 ℃ u nder low and high temperature respectively, and the average development temperat ure and effective accumulated temperature were (557.76?198.95) DD and(236 .02 ?68.20) DD.Conclusion The suitable developme nt temperature of eggs is 27 ℃, the development speed and effective accumulated t e mperature of eggs increases with the temperature under given condition, but too high temperature may have negative impact on the development of eggs. The resul t can be considered as the basic biological parameter of the snail.

Objective: To investigate the use of new types of drugs among HIV/AIDS patients in Ningbo City, Zhejiang Province, so as to provide insights into surveillance and interventions for use of new types of drugs among HIV/AIDS patients. Methods: The HIV/AIDS patients in Ningbo City were randomly sampled from the HIV/AIDS Prevention and Control Information System of Chinese Disease Prevention and Control Information System. Patients' demographics, use of new types of drugs and sexual behaviors were collected using questionnaire surveys. Patients' hair samples were collected, and new types of drugs were determined in hair using liquid chromatography-mass spectrometry (LC-MS). In addition, factors affecting the use of new types of drugs were identified using a multivariable logistic regression model. Results: A total of 254 HIV/AIDS patients were enrolled, including 214 men (84.25%), 31 cases aged under 25 years (12.20%), 66 cases aged 25 to 30 years (25.98%), and 157 cases aged 31 to 75 years (61.81%). There were 30 cases reported previous use of new drugs (11.81%), including 27 cases reported previous use of methamphetamine (90.00%). There were 48 cases tested positive of new types of drugs, with a positive rate of 18.90%, including 44 cases tested positive for methamphetamine (91.67%). Multivariable logistic regression analysis identified age (25 to 30 years, OR=6.926, 95%CI: 1.412-33.969), occupation (students/teachers/cadres/employees/retirees, OR=6.971, 95%CI: 2.123-22.889; housekeeping and housework servants/unemployed, OR=3.356, 95%CI: 1.289-8.739; business/public place servants, OR=2.447, 95%CI: 1.033-4.448) and syphilis infection during recent six months (OR=3.491, 95%CI: 1.664-7.326) as factors affecting the use of new types drugs among HIV/AIDS patients. Conclusions The use of new types of drugs exists in HIV/AIDS patients in Ningbo City. Age, occupation and syphilis infection were associated with the use of new types of drugs among HIV/AIDS patients.

Animals ; Burns ; physiopathology ; Female ; Glycoproteins ; genetics ; metabolism ; Hydrogen Peroxide ; metabolism ; Male ; Mice ; RNA, Messenger ; genetics ; metabolism ; Skin ; injuries ; metabolism ; Wound Healing ; physiology

Objective:To observe the influence of pressing force and time on the thermal effect of An-pressing manipulation.Methods:Eight healthy volunteers were recruited to receive An-pressing manipulation at Xinshu (BL 15) on the right side.The pressing force and time were both divided into five levels:the force described as extremely mild,mild,moderate,strong and extremely strong and time given by 2.5 min,5.0 min,7.5 min,10.0 min and 15.0 min.The real-time change in local acupoint temperature as well as the change during 1.0-15.0 min after the manipulation were observed.Results:Compared with the baseline data,the real-time changes in the temperature after An-pressing Xinshu (BL 15) on the right side with different levels of force (from mild to strong) were respectively (1.88t0.64) ℃,(2.05±0.68) ℃,(2.25±0.59) ℃,(2.35±0.61) ℃ and (2.32±0.69) ℃;the changes in 15.0 min after the manipulation were respectively (-0.11±0.11) ℃,(0.03±0.14) ℃,(0.59±0.58) ℃,(1.38±0.70) ℃ and (2.09±0.98) ℃.The real-time temperature changes after the manipulation for different durations (from short to long) were respectively (1.94±0.37) ℃,(2.33±0.29) ℃,(2.49±0.31) ℃,(2.51±0.39) ℃ and (2.41±0.55) ℃;the changes in 15.0 min after the manipulation were respectively (0.53±0.49) ℃,(0.33±0.30) ℃,(0.52±0.33) ℃,(0.55±0.38) ℃ and (0.76±0.36) ℃.Conclusion:The thermal effect presented an increasing tendency with the extension of pressing time,and the temperature reached the top at 7.5 min;the thermal effect showed an increasing tendency with the rise of pressing force,and the temperature reached the top upon a moderate level of force.The pressing time can produce a greater influence on the real-time temperature than the pressing force;the pressing force can produce a greater influence on maintaining the temperature than the pressing time.

OBJECTIVETo study the clinical pathologic and immunohistochemical features of gastrointestinal mesenchymal tumors (GIMTs), and to investigate the value of molecular markers in GIMTs clinical differentiation diagnosis.

METHODSThe clinical and pathological data of 210 cases of GIMTs, collected from Jan. 1987 to Dec. 2005 in our hospital, were investigated retrospectively. GIMTs were rediagnosed by using standard immunostaining technique in paraffin-embedded tissue. The expression level of CD117, CD34, Desmin, SMA and PS100 were detected by immunohistochemical method.

RESULTSAmong 210 cases of GIMTs, 127 cases were Gastrointestinal stromal tumors (GISTs) (60.5%), 33 leiomyomas and leiomyosarcomas (15.7%), 27 neurogenic tumours (12.8%), and 23 miscellaneous tumors (11.0%). The incidences of GIST, leiomyoma and leiomyosarcoma were similar among men and women. Men were more likely to develop neurogenic tumors and miscellaneous tumors than women. Of all the GISTs, 51.2% cases originated from stomach, 19.7% from small intestine, 11.0% from esophagus, 10.2% from colon and rectum. The most common location of leiomyomas and leiomyosarcomas was esophagus (45.5%). The most common location of neurogenic tumors was retroperitoneum (74.1%). Common symptoms of GISTs included digestive tract hemorrhage in 36 cases (28.3%), abdominal pain in 27 cases (21.3%) and abdominal mass in 24 cases (18.9%). Other GIMT cases except GISTs had no first symptom of digestive tract hemorrhage. It was noticed that 79.5% of GISTs had no obvious invasion, and 72.7% of leiomyomas and leiomyosarcomas had no obvious invasion. 33.3% of neurogenic tumors invaded the adjacent organs or tissues. No metastases had been found in other GIMT cases except GISTs. The neoplastic cells of GISTs were composed of various percentage of spindle (72.5%), epithelioid (11.8%) and mixed-type cells (15.7%). The percentage of spindle cells in leiomyomas and leiomyosarcomas was 94. The immunohistochemical results of GISTs showed that the positive rate of CD117 was 93.7%, CD34 was 69.3%, Desmin was 13.4%, SMA was 12.6%, and PS100 was 10.2%. The immunohistochemical results of leiomyomas and leiomyosarcomas showed that the positive rate of Desmin was 78.5%, SMA was 63.6%, while as the expressions of CD117, CD34, and PS100 were negative. Diffuse strong positive staining of PS100 was observed in 88.9% of neurogenic tumor patients.

CONCLUSIONSGISTs are the most common tumors among GIMTs. GISTs are different from neurogenic tumors, leiomyomas and leiomyosarcomas in initial symptom, tumor location, biological behavior and immunophenotype. Immunohistochemistry plays an important role in differentiating GISTs from leiomyomas and neurogenic tumors.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers ; analysis ; Child ; Child, Preschool ; Female ; Gastrointestinal Neoplasms ; pathology ; Humans ; Immunohistochemistry ; Infant ; Male ; Mesenchymoma ; pathology ; Middle Aged ; Young Adult

OBJECTIVETo study serum concentration and mRNA expression of interleukin-13 (IL-13) in children with steroid-responsive nephrotic syndrome (SRNS) and the effect of methylprednisolone pulse therapy (MPT) on IL-13 expression.

METHODSTwenty-eight children with SRNS were enrolled in this study. Serum protein level of IL-13 was measured using ELISA and IL-13 mRNA expression in peripheral blood mononuclear cells (PBMC) was detected with RT-PCR before MPT, 2 and 5 days after MPT, and 2 weeks after disappearance of proteinuria following MPT. Twenty-four urinary protein was measured with the biuret assay. Twenty healthy children were used as controls.

RESULTSSerum IL-13 levels (38.48 +/- 13.01 pg/mL vs 5.18 +/- 2.71 pg/mL) and PBMC IL-13 mRNA expression (1.31 +/- 0.23 vs 0.36 +/- 0.07) before MPT in SRNS patients were significantly higher than in the controls. After 5 days of MPT and 2 weeks after disappearance of proteinuria following MPT, serum IL-13 levels (15.33 +/- 7.81 and 5.35 +/- 2.12 pg/mL respectively) and PBMC IL-13 mRNA expression (0.89 +/- 0.26 and 0.33 +/- 0.08 respectively) were significantly reduced (P < 0.01). Serum IL-13 levels and PBMC IL-13 mRNA expression in SRNS patients 2 weeks after disappearance of proteinuria following MPT were reduced to control levels, but remained at a higher level than controls 5 days after MPT. A positive correlation was found between serum levels of IL-13 and 24-hour urinary protein in SRNS patients before (r=0.75, P < 0.01) and after 2 and 5 days of MPT (r=0.68, r=0.71 respectively; P < 0.05).

CONCLUSIONSSerum IL-13 levels and PBMC IL-13 mRNA expression in children with SRNS increase. MPT can inhibit the expression of protein and mRNA of IL-13 in these patients.

Adolescent ; Child ; Female ; Humans ; Interleukin-13 ; blood ; genetics ; Male ; Methylprednisolone ; administration & dosage ; Nephrotic Syndrome ; blood ; drug therapy ; Proteinuria ; drug therapy ; RNA, Messenger ; analysis