BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

Objective We investigated the in vivo effects of recombinant adenovirus-associated virus type-2(AAV-2)mediated interleukin-10(IL-10)gene transfer on the expression of matrix metalloproteinase(MMP)-2,9,tissue inhibitor of metalloproteinase(TIMP)-1,collagen type Ⅰ and type Ⅲ in a rat acute myocardial infarction model.Method Male Sprague-Dawley(SD)rats were randomly divided into three groups(each n=6):sham operation group,MI/AAV2 group,and MI/AAV2-IL-10 group(1010 vg/ml×0.1 ml injection at peri-infarct regions immediately post MI).Five days later,the expressions of MMP-2 and MMP-9 were measured by RT-PCR, Western blot and zymography.The expression of TIMP-1 was measured by RT-PCR and Western blot.Collagen type Ⅰ and type Ⅲ were assessed bv RT-PCR and immunohistochemical stain.Results The myocardial expressions of MMP-2,MMP-9 and collagen contents in MI/AAV2 group were significantly increased than those in sham operation group.Myocardial expressions of MMP-2,MMP-9 were significantly decreased and the expression of TIMP-1 significantly increased in the MI/AAV2-IL-10 group than those in MI/AAV2 group. Moreover,the expressions of collagen type Ⅰ,collagen type Ⅲ and the ratio of Ⅰ/Ⅲ collagen in border zones of infarcted myocardium were decreased by 47.6%(P＜0.01),23.6%(P＜0.05),and 17.9%(P＜0.05)respectively,while the expression of TIMP-1 increased by 73.1%(P＜0.05)in MI/AAV2-IL-10 group compared to MI/AAV2 group.Conclusion In vivo myocardial IL-10 transfer reduced myocardial MMP and collagen expression and increasing the TIMP expression.

To establish a method for detecting microdialysis recovery of tetramethylpyrazine (TMP) and ferulic acid (FA) and investigating the influencing factors, providing the basis for further in vivo microdialysis experiments. The concentration of FA and TMP in dialysates were determined by high pressure liquid chromatography ( HPLC) and probe recovery were calculated respectively. The influence of the flow rates, medium concentration, temperature and in vivo probe stability on the recovery of FA and TMP were investigated by using concentration difference method (incremental method and decrement method). The recovery obtained by incremental method were similar to by decrement method. The in vitro recovery rate of FA and TMP decreased with the increase of 1-2.5 μL min(-1), and increased obviously with the temperature of 25-42 degrees C under the same conditions. The concentration of FA and TMP had no obvious effect on the probe recovery under the same flow rate. In addition, the recovery of TMP and FA remained stable and showed similar trends under the condition of four concentration cycles, indicating that the intra day reproducibility of the concentration difference method was good. The recovery of brain microdialysis probes in vivo 8 h maintained a relatively stable, but certain differences existed between different brain microdialysis probes, demonstrating that each probe was required for recovery correction in vivo experiment. Microdialysis sampling can be used for the local brain pharmacokinetic study of FA and TMP, and retrodialysis method can be used in probe recovery of FA and TMP in vivo.
Animals ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; Coumaric Acids ; analysis ; isolation & purification ; pharmacokinetics ; Drugs, Chinese Herbal ; Humans ; Microdialysis ; methods ; Pyrazines ; analysis ; isolation & purification ; pharmacokinetics ; Rats

OBJECTIVESTo investigate incidence of perioperative cardiovascular events, to analyze related risk factors for the patients undergoing intraperitoneal surgery.

METHODSThe data of 1079 patients who underwent intraperitoneal surgery (exclude laparoscope surgery) from July 2007 to June 2008 was reviewed and analyzed.

RESULTSFor the patients undergoing intraperitoneal surgery, the incidence of major cardiovascular events was 3.99% (43/1079), all-cause mortality was 1.58% (17/1079). The independent risk factors of major cardiovascular events were age ≥ 60 years, history of coronary heart disease, cardiac insufficiency, arrhythmia, chronic obstructive pulmonary disease, estimated glomerular filtration rate (eGFR) < 60 ml/(min·1.73 m(2)), emergency surgery and duration of surgery > 2.82 h (OR = 2.68 to 5.19, P = 0.001 to 0.031).

CONCLUSIONSThe cardiac risk of intraperitoneal surgery is 3.99%. The risk of cardiac complications should be evaluated in elderly patients and those with ischaemic heart disease, chronic obstructive pulmonary disease, and renal disease, more specifically, when emergent or long duration major surgeries are needed.

Abdomen ; surgery ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cardiovascular Diseases ; epidemiology ; mortality ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; mortality ; Risk Factors ; Young Adult

OBJECTIVETo design and manufacture a reliable spine phantom used in the cross calibration and quality control of dual X-ray absorptiometry (DXA).

METHODSA hydroxyapatite quality control phantom was designed and made through three steps: solid water, bone phantom material and integration, then evaluated the phantom on four different types of DXA machines made by LUNAR company.

RESULTSAmong the four DXA densitometers, the Expert fan beam densitometer had the biggest accuracy errors of bone mineral density (BMD), bone mineral content(BMC) and area values while the other three one narrow fan beam (Prodigy) and two pencil beam densitometers-had small errors. Of the three indexes measured by all the machines, BMD error was the smallest (-15.4%-11.5%), with the Prodigy's BMD was most outstanding. BMD errors at the higher density ends were small, tend to be positive values while the errors at the lower ends were big, tend to be negative. In cross calibration, giving consideration to the differences between the both ends, it is better to use the regression equation to correct. The base line of precision error derived by scanning the phantom once a day for consecutive 25 days was better than that derived by scanning the phantom 25 times consecutively on the same day. As to precision error, the coefficient of variation (CV) of scanning-25 times-a day was the smallest (0.0043) while the CV of 12 adults measurements was the biggest (0.0078).

CONCLUSIONSThis phantom can be used in the quality control and cross-calibration of different types of DXA machines.

Absorptiometry, Photon ; Adolescent ; Adult ; Biocompatible Materials ; Bone Density ; Calibration ; Durapatite ; Female ; Humans ; Male ; Middle Aged ; Models, Anatomic ; Spine

A sensitive high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method was established for the determination of eplerenone (EP) in human plasma. The plasma samples of EP were extracted with ethyl acetate and separated by HPLC on a reversed phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate water solution-methanol (30 : 70, v/v). EP was determined with electrospray ionization-mass spectrometry (ESI-MS) in the selected ion monitoring (SIM) mode. The calibration curves were linear over the range of 2-4 000 ng x mL(-1) for EP. The lower limit of quantification was 2 ng x mL(-1). The method has been successfully applied in the pharmacokinetic study of the EP tablets. The main pharmacokinetic parameters of EP after oral administration of 25 mg, 50 mg, 100 mg were as follows, t1/2: (4.9 +/- 2.1), (4.7 +/- 1.5), (5.9 +/- 1.2) h; AUC(0-infinity): (4 402 +/- 1 735), (8 150 +/- 2 509), (13 783 +/- 4 102) microg x h x L(-1); and MRT: (6.2 +/- 2.1), (6.6 +/- 1.3), and (7.2 +/- 1.6) h. Parameters of EP after oral administration of multiple doses of 50 mg were as follows, t1/2: (6.1 +/- 1.7) h; AUC(ss): (10 071 +/- 4220) microg x h x L(-1); MRT: (8.1 +/- 2.3) h; and DF: (3.2 +/- 1.0).
Chromatography, High Pressure Liquid ; methods ; Humans ; Spectrometry, Mass, Electrospray Ionization ; methods ; Spironolactone ; analogs & derivatives ; blood ; pharmacokinetics

Objective] To study the effect of different placed time for vacutainer and specimen on the results of emergency electrolyte detection . [ Methods] With heparin lithium anticoagulation tube and common coagulation vacuum tube , electrolytes were detected at 30 minutes,one hour and two hours af-ter extracting blood . [ Results] At 30 minutes and one hour after extracting blood ,the levels of K +and Na+of the plasma group were significantly lower than those of the serum group (P<0.05).With the ex-tension of specimen placed time , the levels of K +and Na +of the plasma group were becoming higher than those previously ,and at two hours the difference had statistical significance ( P<0 .05 ) .There was not ob-vious difference found in the levels of K +, Na +and Cl -of the serum group at different placed times ( P>0.05).But the levels of CO2 of both the plasma group and the serum group were significantly lower than those previously with the extension of specimen placed time , and the difference had statistical significance (P<0.05). [Conclusion] It is indicated that the using of different vacutainers effects electrolyte de-tection.With the extension of specimen placed time , the levels of K +and Na +of the plasma group increase gradually , and the levels of CO 2 of the plasma group and the serum group both decrease gradually .

Adult ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; diagnosis ; Humans ; Male ; Middle Aged ; Prognosis

The plaque-forming characteristics of Newcastle disease viruses of chickens and geese source were compared on various cells. The result showed that there were obvious differences of plaque formation between F48E9 and NA-1 on Vero cells, chicken embryo fibroblast cells (CEF) and goose embryo fibroblast cells (GEF). The plaque-forming ability of NA-1 was higher than F48E9 on GEF, but lower than F48E9 on CEF. On Vero cells, the plaque-forming ability of NA-1 was slightly stronger than F48E9. It demonstrated that the plaque-forming characteristics were consistent with host tropism of virus. The morphogenesis of F48E9 and NA-1 on Vero cells was observed with transmission electron microscope. There were different replication processes between F48E9 and NA-1 on cells at different stages. NA-1 had stronger adaptability to host than F48E9 according to budding processes and envelope integrity.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Geese ; Host-Pathogen Interactions ; Newcastle Disease ; virology ; Newcastle disease virus ; growth & development ; isolation & purification ; physiology ; ultrastructure ; Poultry Diseases ; virology ; Vero Cells ; Viral Plaque Assay

AIMThe process of vascular calcification involves various genetic alterations which may play a very important role in the vascular calcification. Vascular smooth muscle cells undoubtedly composed the main part of vascular cells, and are involved in vascular calcification. So bovine artery smooth muscle cell (BASMC) was used to investigate the gene changes during BASMC's calcification.

METHODSBovine artery smooth muscle cells cultured in vitro was induced calcified by beta-Glycerophosphate (beta-GP). Using DD-PCR technique to screening differentially expressed genes and those differentially expressed bands were reexamined by reverse Northern blot. All the ESTs were sequenced and BLAST with GenBank.

RESULTSTotal 65 cDNAs were isolated as differentially expressed genes and 40 of them were successfully reamplified. Using reverse-Northern blot, seven of these 40 cDNAs were reproducibly expressed differentially between the two cells. Three of them are new bands and have not been reported before.

CONCLUSIONThis is the first time using DD-PCR to screen differentially expressed genes of BASMC calcification. Seven related ESTs were identified relating to BASMC calcification.

Animals ; Arteriosclerosis ; genetics ; metabolism ; pathology ; Cattle ; Cells, Cultured ; Expressed Sequence Tags ; Genetic Variation ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Vascular Calcification ; genetics ; metabolism ; pathology