Objective To study the main subtypes messenger ribonucleic acid(mRNA) and the basal enzyme activity of phosphodiesterase (PDE) in rat pulmonary microvascular endothelial cells (PMVECs) through the examination mRNA expression and activity of PDE in vitro. The data were offered to reveal the relationship between PDE distributions, activity change and to accumulate data for the possibility of drug regulation of its functional alteration. Methods The cells were cultured with tissue-sticking method;the gene expression of PDEs was detected by reverse transcript polymerase chain reaction (RT-PCR), and the activity of PDEs was calculated by cyclic nucleotides content change examined with high performance liquid chromatogram (HPLC) before and after the PDE reaction( n ＝3). Results The PMVECs identified by cell immunofluorescence with polyclonal antibody of CD31 were dissociated and cultured, mRNAs of PDE1A, 1C, 2A,3A, 3B, 4A, 4D, 5A, 7A, 7B, 8A, 8B, 9A, 10A,11A were expressed in PMVECs, but there was no mRNA of PDE1B expressed in PMVECs. cAMP/cGMP-PDE in the extent of 5-20μl had a good linear correlation with its activity. Conclusion There are 17 kinds of PDE gene expression existing in PMVECs which contain of the basic enzyme with a higher activity.

Objective To assess the clinical value of laparoscopic colorectal resection combined with simultaneous radiofrequency ablation for liver metastases.Methods Twenty-two patients with colorectal cancer and synchronous liver metastases were treated by laparoscopie colorectal resection and simultaneous radiofrequency ablation(RFA)in Third People's Hospital of Chengdu from December 2001 to July 2006.Areas of colliquative necrosis were detected by contrast-enhanced computed tomography postoperatively.The outcomes were analyzed by chi-square test.Results Of all patients,8 had muhiple liver metastases,16 had complications.Thirty-one liver metastases were treated by RFA,and no complication oecurred.The mean postoperative hospital stay was(14±5)days,and there was no perioperative death.Five of the 22 patients were identified as incomplete ablation of the tumor,and the RFA was reperformed.The recurrence of liver metastases in the RFA necrotic zone was observed in 4 patients,and RFA was reperformed on 2 of them.Six patients died,and 2 of them died of liver metastases recurrence in the RFA necrotic zone.The recurrence rate of liver metastases and mortality was 18%(4/22)and 27%(6/22),respectively.The recurrence rate of patients with the diameter of the metastatic lession≥2.0 cm was significantly higher than those with the diameter of the metastatic lession＜2.0 cm(x2=5.867,P＜0.05).Conclusions Laparoseopie colorectal resection combined with simultaneous RFA provide a curative opportunity for colorectal cancer patients who are old and have multiple liver metastases,complications,poor tolerance of operation,and for patients whose tumors are difficult to resect.

Objective To explore the different apoptotic gene expressions and apoptotic signaling transduction of human rhabdomyosarcoma (RD) cells infected by enterovirus 71 (EV71) in different stage.Methods The survival of EV71-infected RD cells was observed by trypan blue staining.The apoptotic morphology and rates of RD cells were surveyed and detected by Annexin V-FITC/PI staining and flow cytometry,respectively.PCR array was employed to analyze 88 apoptotic gene expressions from EV71-infected RD cells at 8 h and 20 h postinfection (p.i),respectively.Results After RD cells were infected with EV71 (MOI =5) at 8 h p.i,the viability was significantly decreased.Flow cytometry data demonstrated that the apoptotic rates of EV71-infected RD cells had increased to 18.0％ and 19.1％ at 20 h p.i in early and later stage respectively.RT-PCR array studies revealed significant variations in the expression of apoptotic genes.Among 88 genes,only the expression of IFN-β1 was upregulated 5.22 folds,whereas 47 genes including ACIN1,Akt,Apaf1,caspase and CIDEB were found to be downregulated that were lower than 2 folds at 8 h p.i.However,28 genes including FasL,CD40L,TNF,caspase-10 and caspase-3 were induced more than 2 folds after EV71 infection at 20 h.Conclusion The downregulation of apoptosis-related genes is associated with viral apoptosis-suppressing effect in RD cells in the early stage of EV71 infection.The death receptor signaling pathways including Fas/FasL and TNF/TNFR are activated to induce cell apoptosis in the late stage of EV71 infection.Moreover,host cell can also inhibit apoptosis by regulating signal pathway of CD40/CD40L,NF-κB/RelA and PI3K/Akt activation.

Objective To study the correlation between morphological characteristics of coronary artery calcified plaques and luminal stenosis of local coronary artery segments with 64-slice computed tomography.Methods One hundred and eleven patients who had undergone 64-slice computed tomographic coronary angiography(CTCA)and conventional coronary angiography(CAG)were retrospectively analyzed.The calcified plaques were classified as punctate,nodular,strip-like and nubbly in long-axis view of coronary artery lumen,and were classified as crescent,semilunar,round moon and circinate in short-axis view.The morphologic characteristics of these calcified plaques on CTCA were retrospectively analyzed and compared with luminal stenosis of CAG results.Results Among 528 calcified coronary segments which were analyzed in 111 patients,there were 383(72.5%)punctate calcified plaque segments and 145(27.5%)of non-punctate plaques.There were 34(23.4%,34/145)non-punctate calcified plaques which caused severe stenosis(≥75%),including 4(11.8%)nodular,8(23.5%)stripe-like and 22(64.7%)nubbly calcified plaques on the long-axis view,and 0(0.0%)cresent,8(23.5%)semilunar,18(52.9%)round moon and 8(23.5%)circinate calcified plaques on the short-axis view.The ratios of different morphological coronary artery calcifications which caused severe stenoses were significantly different with each other(all P＜0.01).Conclusion Different figures of coronary artery calcified plaques demonstrate different degrees of stenoses of local coronary artery lumen.Severe stenoses were mostly caused by nubbly calcified plaque on long-axis view,round moon and circinate calcified plaque on short-axis view.

BACKGROUNDFolic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of folic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase (DHFR) gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 (HAND2) and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations.

METHODSMorpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression.

RESULTSDHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos (P < 0.05). Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos.

CONCLUSIONSDHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; physiology ; Female ; Heart ; embryology ; Heart Defects, Congenital ; etiology ; Tetrahydrofolate Dehydrogenase ; genetics ; physiology ; Zebrafish ; Zebrafish Proteins ; genetics ; physiology

OBJECTIVETo study the preparation of liposomes surface-modified with glycyrrhetinic acid targeting to hepatocytes.

METHOD3-succinic-30-stearyl glycyrrhetinic acid(Suc-GAOSt), one of the amphiphilic glycyrrhetinic acid derivatives, was synthesized as targeting molecules, liposomes surface-modified with glycyrrhetinic acid has been produced with ethanol injection method.

RESULTTargeting molecules can be mixed into the liposomal membrane. It was confirmed that the targeting molecules is 9% of the total lipids at the most in the liposomes.

CONCLUSIONLiposomes surface-modified with glycyrrhetinic acid was successfully prepared, which is considered to be a potential approach targeting to hepatocytes.

Drug Carriers ; Drug Delivery Systems ; methods ; Glycyrrhetinic Acid ; administration & dosage ; Hepatocytes ; Liposomes ; Particle Size ; Phospholipids ; Succinic Anhydrides

AIMTo study the preparation of bovine serum albumin nanoparticles surface-modified with glycyrrhizin(BSA-NP-GL) targeting to hepatocytes.

METHODSThe bovine serum albumin nanoparticles (BSA-NP) were prepared by desolvation process. Glycyrrhizin (GL) was oxidized by sodium periodate to be conjugated to surface reactive amino groups (SRAG) of the BSA-NP. The SRAG were quantified by spectrophotometric method using 2, 4, 6-trinitrobenzenesulfonic acid(TNBS). Glycyrrhetinic acid(GA) hydrolyzed from GL, which was on the surface of BSA-NP-GL was assayed by HPLC after isolation by sephadex G-50. Both methods were used to verify the conjugation achieved. HPLC was used to determine surface density of GL on BSA-NP-GL.

RESULTSThe amount of SRAG of the BSA-NP-GL decreased by 19.6% compared with normal BSA-NP. The amount of GL molecule was 9.2% of the total determined SRAG of BSA-NP. The mean diameter of the BSA-NP-GL was 73 nm with round shape. The stability of BSA-NP-GL was constant when it was stored at 25 degrees C and 37 degrees C during 10 days.

CONCLUSIONBSA-NP-GL was successfully prepared, which is considered to establish an experimental foundation for further research on its ability for targeting to hepatocytes.

Cross-Linking Reagents ; chemistry ; Drug Delivery Systems ; Glycyrrhizic Acid ; chemistry ; Nanotechnology ; Particle Size ; Serum Albumin, Bovine ; administration & dosage ; chemistry ; ultrastructure ; Surface Properties ; Technology, Pharmaceutical ; methods

A cross-sectional survey with multiple-stage and random sampling was performed among middle and elderly aged permanent inhabitants in Wuhan area.The prevalences of metabolic syndrome,diabetes mellitus, impaired glucose tolerance,hypertension and obesity were 12.2%,11.8%,10.3%,31.9% and 48.0% respectively.

Objective To analyze the image quality of coronary arteries by dual-source computed tomography (DSCT) in children with congenital heart disease(CHD). Methods Image quality of coronary arteries were scored by two radiologists with an ordinal grading system. Score 3 or 4 was considered to be good image quality and diagnostic. Score differences among 3 groups of various baseline heart rate levels, 3 groups of various age levels and 4 coronary branches were analyzed by using ANOVA. Multiple linear regression analysis was used to test the effects of age, heart rate, tube voltage, tube current, pitch, contrast volume, contrast injection rate on the coronary image scores. Results A total of 72 consecutive patients were enrolled. 71.2% (205/288) of coronary artery branches was good enough for diagnosis. Image quality of left main coronary artery (LM, score 3.64±0.49), left anterior descending (LAD, score 2.97±0.40), right coronary artery (RCA, score of 2.82±0.70), left circumflex (LCX, score 2.76±0.73) were in a descending order (F=29.00, P<0.01). Image scores of age groups of 0--1 years old (2.80±0.65), >1--3 years old (2.81±0.74), >3--14 years old (3.58±0.52) were in an ascending order (F= 20.16,P<0.01). Image scores of heart rate groups of<100 bpm (3.54±0.56),100--150 bpm (2.86± 0.70) and >150 bpm (2.81±0.63) were in a descending order (F=16.72, P<0.01). Multiple linearregression analysis demonstrated that multiple impact factors affected image quality of LM, LAD, LCX and RCA. Age(r=0.013 ,P<0.01)and contrast volume(r=0.292, P<0.01)had positive correlation with LM score. Age(r=0.509, P<0.01) and voltage (r=0.292, P=0.011) had positive correlation with LAD score. Heart rate(r= -0.179, P<0.05) had negative correlation with LAD score. Age (r=0.063, P< 0.01) had negative correlation with LCX score. Age (r=0.486,P<0.01) and voltage (r=0.220, P<0.05) had positive correlation with RCA score. Conclusions Dual-source CT could clearly shows coronary arteries of children with CHD over the age of 3 or with heart rate less than 100 bpm. Limitations of Dual-source CT coronary angiography for CHD children included young age and fast heart rate.

Objective To evaluate the value of MSCT, MRI and SPECT in detecting acute myocardial ischemia in Chinese mini-swine model. Methods A total of six male mini-pigs were recruited with a mean body weight of (21.6 ± 1.2) kg. All pigs were scanned on MSCT before the ligation of distal segment of Left anterior descending artery. Then, MSCT was rescanned every 2 h from ligation till 8 h latter.MRI, SPECT and the last MSCT scan were performed within 24 h one by one. Finally pathological examination was carried out right after the pig killed. Results One pig died during operation, the other 5 finished all the examinations. The pathological staining showed the same areas of myocardial infarction in the left ventricular anterior wall with the all the imaging findings, including low perfusion region of MSCT arterial phase at 2-24 h, low perfusion region of SPECT at 24 h and low perfusion region of MRI first pass phase at 24 h. Three of 5 pigs showed enhanced edge of low perfusion region on MSCT delayed scan at 4-8 h. The mean CT values in the region with reduced first-pass perfusion were 75.9,36.4, 35. 2,37. 8,37.4,33.3 HU on MDCT image at baseline, 2-8 h after operation and within 24 h after operation, respectively,and there were statistically significant difference of CT values ( F = 12. 341, P ＜0. 01 ) between preoperative and all postoperative MSCT scan. There were no statistically significant difference (F = 2. 278, P = 0. 792)among all postoperative MSCT scan. At baseline, 2-8 h after operation and within 24 h, the average volumes of stroke volume(SV)were 21.7,11.9,10.3,11.4,12. 3,12.6 ml, respectively, while the average volume of end-systolic volume( ESV)were 15.2,23.4,25.0,24. 4,25.3,22. 8 ml,respectively. The average volume of end-diastolic volume ( EDV ) at these time point were 37. 0,35.4,35.0,35.7,37. 6,37.5 ml,respectively and the average percentage of ejection fraction (EF) were 58.9% ,33.8% ,29. 0%, 31.9%,32.6% ,33.5% ,respectively. SV(F =22. 349, P＜0.01) ,ESV (F=8. 810, P＜0.01) ,EF(F =27. 240,P ＜ 0. 01 ) were all significantly different among all postoperative MSCT scan except EDV ( F = 2. 339, P =0. 079). Infarct size, which was defined as the proportion of the area of infarction to that of the entire heart,were (39.4 ±12.6)% for MSCT,(37.2 ± 10.0)% for MRI, (35.9 ±9.6)% for TTC, respectively.There were no significant differences of infarct size between TTC and MSCT (t =0. 612, P =0. 574), TTC and MRI (t=0.820, P=0.458), MSCTand MR (t=0. 425 ,P =0. 692 ). Conclusions MSCT,MRI and SPECT were all able to be used to detect the myocardial infarction in acute myocardial ischemia model The infarct size defined on MSCT, MRI and pathology were consistent. The density of ischemic myocardium and cardiac function did not change over the time within 24 h right after infarction.