Objective To investigate the inhibition of low-dose aprotinin on systematic inflammatory response during cardiopulmonary bypass. Methods Twenty-eight ASA Ⅱ-Ⅲ patients(male 13, female 15) undergoing valve replacement were studied. The age ranged from 27 to 55 years and body weight from 37 to 70 kg. CPB time ranged from 66 to 218 mm and aortic cross clamping time from 30 to 140 min . The patients were divided into two groups: control group(n=14) and aprotinin group(n = 14) Premedication included intramuscular phenobarbital sodium 0.1 g and scopolamine 0.3 mg. Anesthesia was induced with intravenous midazolam 0. lmg/kg, fentanyl 20-30?g/kg and vecuronium 0. 1-0. 12mg/kg, and maintained with intermittent bolus injection of fentanyl and vecuronium supplemented with isoflurane inhalation. In aprotinin group aprotinin 1?106KIU was infused after induction of anesthesia until thoracotomy, after thoracotomy aprotinin was infused at a rate of 2.5? 106 KIU/h and aprotinin 2. 5 ?106KIU was added to the prime fluid as suggested by Levy. Blood samples were obtained before anesthesia, before CPB and 1h and 24h after CPB for determination of CD11b and CD18 expression on the surface of neutrophil by immunofluoresence flow cytometry. Results The expression of CD11b or CD18 and CPB time were positively correlated(?= 0.644, 0.538, P0.05). 1 h after CPB CD11b/ CD18 expression increased significantly in the control group and was significantly higher than that in the aprotinin group, but at 24h after CPB only CD11b expression in control group was significantly higher than that in aprotinin group. In aprotinin group there was no significant difference in CD11b/CD18 expression between the four intervals.Conclusions CPB-induced systematic inflammatory response may be positively correlated with CPB time. Low dose aprotinin can inhibit the increase in CD11b/CD18 expression after CPB.

Objective To evaluate the roles of magnetic resonance imaging and proton magnetic resonance spectroscopy(1H-MRS) in the diagnosis of meningiomas. Methods 98 patients with meningiomas underwent conventional pre-contrast MR and contrast MR. Among them, 28 cases had two dimensional single voxel or multi voxel 1 H-MRS simultaneously both in the lesion's region and the contralateral side. Results On precontrast MR images of 98 cases, T1 WI showed 58.1% (61/105) isointensities, 31.4% (33/105) faintly low intensities and 10. 5% (11/105) mixed intensities; T2WI showed 40. 0% (42/105) isointensities, 41.0%(43/105) hyperintensities, 10.5% (11/105) faintly low intensities and 8.5% (9/105) mixed intensities. After administration of Gd-DTPA, the solid part of the tumors exhibited various enhancement in all the 98 cases.28 cases of MRS exhibited specific different spectral peaks, including increased of choline-containing compounds(Cho), absent or decreased of acetylaspartate(NAA), and the unchanged of creatine(Cr). The value of NAA, Cr, Cho, NAA/Cr, Cho/Cr, NAA/Cho in the tumor center of meningioma were 0. 09 ± 0.06,0.31 ± 0. 22, 0.46 ± 0. 16, 0.33 ± 0. 42, 1.50 ± 0. 68, 0. 15 ± 0.08, compared with the contralateral normal region, Cr has no significant difference (P ＞ 0. 05), NAA, Cho, NAA/Cr, Cho/Cr, NAA/Cho had significantly differences(P ＜ 0.05). Conclusion Conventional pre-contrast MR and contrast MR is the most important dignostic means for meningiomas, 1H-MRS combined with MRI can improve the diagnostic accuracy of meningiomas.

In this paper, five different methods were carried out for DNA extraction directly from soil. The result shows that all five methods could generate DNA with more than 15 kb in size. They were subsequently used as templates for PCR amplification with success, using primers of the bacterial 16S rRNA gene and Shiva-1 gene encoding an antibacterial peptide. However, method 5 is more suitable for DNA extraction directly from a small amount of soil sample as it produced a good yield of DNA in high integrity with reliable reproducibility.

Objective To localize the sinoatrial node (SAN) of the newborn rabbits in vivo and cut it for purifying cultivation and study the morophologic characters of primary cultured pacemaker cells of SAN under light microscope and transmissional electron microscope. Methods Hearts of the newborn rabbits were embedded in paraffin for HE-staining and observed the location, form of SAN under optical microscope; SAN cells isolated from neonatal rabbits cultured and purified with the method of differential attachment and BrdU-treatment.Results SAN localized in the anterior wall of the superior vena cava and the posterior-lateral atrial wall.There was about 0.32 mm between its lowest point and sulcus terminalis. Three distinctly different types of cells were observed among the cultured cells of SAN: spindle, araneiform and polygon. The spindle cells covered the greatest proportion of the cultured cells of SAN (59.6%?7.3%). The frequency of spontaneous contraction of spindle cells was the highest among the constrcting cells (145 ?9)time/min. The results of ultrastructure observation showed that myofibrils and other organelles in spindle cells were poorly organized and significantly decreased in number compared with araneiform cells. There was no significant difference between araneiform cells isolated from SAN and from atrial muscle.Conclusion Among the cultured cells from neonatal rabbits SAN, the spindle cells are the pacemaker cells of SAN.

OBJECTIVETo explore etiology, clinical manifestation and immunological changes of infectious pneumonia of neonates in Chengdu area.

METHODSSerum specimens were collected from 111 infants with infectious pneumonia. Eight viral and mycoplasmal specific serum IgM antibodies were detected by enzyme linked immunosorbent assay (ELISA); C reactive protein (CRP), total IgG and its subclasses, IgA and IgM were determined by rate scattered nephelometry; T lymphocyte subpopulations were detected by biotin-streptavidin-peroxidase method, and clinical and other laboratory data were analyzed.

RESULTS(1) Etiological agents: specific serum IgM antibodies were positive in 40 of 111 cases (36.0%) with pneumonias. All the 30 control infants were negative for the specific serum IgM antibodies. Among 111 infants with infectious pneumonia, 20.7% had single viral or mycoplasmal infection, 40.5% had bacterial infection, 15.3% had viral and mycoplasmal infection with bacterial infection; 23.4% had infection with unknown agents. (2) The most common clinical manifestations were tachypnea and cyanosis. The next were cough, milk choking, rales, retractions of the supraclavicular, intercostal and subcostal areas. Roentgenographic examination commonly revealed vague opacities, increased density and patchy infiltration. (3) Immune status: (1) CD(3), CD(4) cell counts of infants with pneumonias were lower than those of the controls while their serum IgA, IgM concentrations were higher than those of the control. (2) The CD(3) and CD(4) cell counts of the group with bacterial infection were lower than those of the control group. (3) The serum IgA concentration of the group with viral and mycoplasmal infection was higher than those of the control group and the group with unknown infection. (4) The serum IgM concentration of the group with bacterial infection was higher than those of the control group. (5) There were no significant differences in CD(8) cell counts, CD(4)/CD(8), concentration of serum IgG and IgG(1 - 4) between pneumonia group and the control group, and among various infectious groups and the control.

CONCLUSIONPathogens of neonatal infectious pneumonia in Chengdu area included single viral or mycoplasmic infection or bacterial infection, viral and mycoplasmal infection with bacterial infection, and unknown infection. Immunological changes of newborn infants suffered from infectious pneumonia included declined CD(3) and CD(4) cell counts, particularly in bacterial infection.

Antibodies, Bacterial ; blood ; Antibodies, Viral ; blood ; Bacterial Infections ; complications ; C-Reactive Protein ; analysis ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin M ; blood ; Infant, Newborn ; Male ; Pneumonia ; diagnosis ; etiology ; immunology ; T-Lymphocyte Subsets ; immunology ; metabolism ; Virus Diseases ; complications

According to the super-position principle of the reinforcement of biological activities, a series of novel E-substituted 2, 3-diaryl propenoic acyloxy phosphonate derivatives were designed and synthesized. And the structures of the target compounds were confirmed by IR, 1H NMR, 13C NMR and elemental analysis. Furthermore, the cytotoxicities of all compounds on A-549, SGC-7901 and EC-109 in vitro were evaluated by MTT assay, and some of them showed good antitumor activity. Among the active compounds, especially, the IC50 value of compound 3e was (12.7 ± 1.9) μmol x L(-1) against A-549 cells, similar to cisplatin [IC50 = (8.0 ± 1.5) μmol x L(-1)], compounds 3g and 3k had better inhibition effect on EC-109 cells growth, with the IC50 values of (9.5 ± 1.8) μmol x L(-1) and (11.5 ± 0.9) μmol x L(-1) respectively, and compounds 3i and 3k exhibited good cytotoxic property on A-549, SGC-7901 and EC-109, which were worth further investigation.
Antineoplastic Agents ; chemical synthesis ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Drug Design ; Humans ; Organophosphonates ; chemical synthesis ; pharmacology

BACKGROUNDAscorbic acid has important antioxidant properties, and may play a role in the protective effects of ischemic preconditioning on later ischemia-reperfusion. Herein, we examined the role of endogenous extracellular ascorbic acid in ischemic preconditioning in the kidney.

METHODSWe developed a solitary rabbit kidney model where animals received ischemia-reperfusion only (ischemia-reperfusion group, n = 15) or ischemic preconditioning followed by ischemia-reperfusion (ischemic preconditioning group, n = 15). Ischemia-reperfusion was induced by occluding and loosening of the renal pedicle. The process of ischemic preconditioning included 15-minute brief ischemia and 10-minute reperfusion. In vivo microdialysis coupled with online electrochemical detection was used to determine levels of endogenous extracellular ascorbic acid in both groups. The extent of tissue damage was determined in kidney sections stained with hematoxylin and eosin. Serum creatinine and urea nitrogen were also detected to assess renal function.

RESULTSDuring ischemia-reperfusion, the extracellular ascorbic acid concentration during ischemia increased rapidly to the peak level ((130.01 +/- 9.98)%), and then decreased slowly to near basal levels. Similar changes were observed during reperfusion (peak level, (126.78 +/- 18.24)%). In the ischemic preconditioning group there was a similar pattern of extracellular ascorbic acid concentration during ischemic preconditioning. However, the ascorbic acid level was significantly lower during the ischemia and early reperfusion stage compared to the ischemia-reperfusion group. Additionally, the extent of glomerular ischemic collapse, tubular dilation, tubular denudation, and loss of brush border were markedly attenuated in the ischemic preconditioning group. Levels of serum creatinine and urea nitrogen were also decreased significantly in the ischemic preconditioning group.

CONCLUSIONSIschemic preconditioning may protect renal tissue against ischemia-reperfusion injury via use of extracellular ascorbic acid. In vivo microdialysis coupled with online electrochemical detection is effective for continuous monitoring extracellular ascorbic acid in the renal cortex.

Animals ; Ascorbic Acid ; metabolism ; Disease Models, Animal ; Ischemic Preconditioning ; methods ; Kidney ; metabolism ; pathology ; Rabbits ; Reperfusion Injury ; prevention & control

On the basis of the chromogenic reaction between Hg and CuI, a semi-quantitative solid sampling Hg analyzer comprising the catalytic furance, Hg testing tube, air pump and smart cellphone was developed. White carrier 101 was chosen as the adsorbent for CuI to react with Hg from the catalytic furnace. The established Hg analyzer can not only visually recognize the coloration when Hg exceeding the limit standard, but also semi-quantitatively detect the Hg content in cosmetics fast using a smart cellphone and RGB analysis software, after direct solid sampling introduction of cosmetics sample. The instrumental detection limit ( LOD) of mercury was 50 ng, the linearity ranged from 50 ng to 2500 ng, the linear regression coefficient ( R2) was higher than 0. 97, and the RSD of the corresponding RGB values was 6% ( n=11 ) . Nine real cosmetics samples were measured by the established method, whose relative differences of Hg contents with that by the standard method (Safety Technical Specification for Cosmetics, 2015 edition) were less than 10% . The whole analytical time can be controlled within 5 min. The established instrumental method is simple, fast, accurate and visual, and extremely suitable to fast and on-site monitoring of Hg in cosmetics samples.

The aim of present study was to explore the vasodilatation mechanism of angiotensin II (AngII) at the molecular level by investigating the effect of AngII on large-conductance Ca²⁺-activated potassium channels (BK(Ca)) in human mesenteric artery smooth muscle cells. The effect of AngII on BK(Ca) was observed by using patch clamp single channel recording technique and amphotericin-perforated whole-cell recording technique. AngII type 1 receptor (AT₁R) and AngII type 2 receptor (AT₂R) mRNA expression in human mesenteric artery was detected by RT-PCR. In cell-attached patch (Vm = +40 mV), AngII (100 nmol/L) had no significant effect on BK(Ca). After pretreatment with Valsartan (a specific inhibitor of AT₁R, 10 μmol/L), 25, 100 and 250 nmol/L AngII stimulated BK(Ca) activity significantly in a dose response manner. After pretreatment of Valsartan, AngII (100 nmol/L) enhanced BK(Ca) open probability (NP(O)) from 0.010 ± 0.003 to 0.039 ± 0.015, decreased the mean close time (T(C)) of BK(Ca) markedly from (2 729.5 ± 808.6) ms to (487.7 ± 182.5) ms (n = 11, P < 0.05) , but AngII had no significant influences on the amplitude (Amp) and the mean open time (T(O)) of BK(Ca). Further PD123,319 (a specific inhibitor of AT₂R) treatment prevented the stimulatory effect of AngII: PD123,319 decreased the NP(O) of BK(Ca) from 0.016 ± 0.003 to 0.004 ± 0.001 (n = 5, P < 0.05), but had no significant influences on Amp, T(O) and T(C) of BK(Ca). In addition, after pretreatment with Valsartan and PD123,319, AngII (100 nmol/L) had no significant effect on BK(Ca). In the amphotericin-perforated whole-cell patch-clamp configuration, after pretreatment with Valsartan, the current density of BK(Ca) at the voltage of -60 - +30 mV had no significant changes before and after adding 100 nmol/L AngII, but the current density of BK(Ca) at the voltage of +40 mV, +50 mV and +60 mV increased significantly after adding 100 nmol/L AngII, from (9.03 ± 2.23) pA/pF, (12.88 ± 2.55) pA/pF and (17.26 ± 2.84) pA/pF to (12.47 ± 2.22) pA/pF, (18.71 ± 2.51) pA/pF and (27.21 ± 3.12) pA/pF (n = 6, P < 0.05), respectively. Using RT-PCR, the AT₁R mRNA and AT₂R mRNA from isolated human mesenteric artery were detected. So we can draw a conclusion, AngII can stimulate BK(Ca) activity in human mesenteric artery smooth muscle cells after pretreatment with Valsartan, which is possibly mediated by AT₂R.
Angiotensin II ; pharmacology ; Humans ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Mesenteric Arteries ; cytology ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Patch-Clamp Techniques ; Receptor, Angiotensin, Type 1 ; metabolism ; Receptor, Angiotensin, Type 2 ; metabolism ; Tetrazoles ; pharmacology ; Valine ; analogs & derivatives ; pharmacology ; Valsartan ; Vasodilation

OBJECTIVETo investigate the signal transduction mechanism underlying the effects of angiotensin II (AngII) on extracellular signal-regulated kinase 1/2 (ERK1/2), early growth response-1 (EGR-1) and platelet-derived growth factor-B (PDGF-B) in hepatic stellate cells (HSCs).

METHODSHSC-T6 cells treated with AngII for 10 or 30 min were examined for phospho-P42/44 protein expression using Western blotting. In another experiment, the cells were preincubated for 1 h in the presence of U0126 (an inhibitor of the MAPK/ERK kinase), irbesartan (an AT-1 receptor blocker), or antioxidant-N-acetylcysteine (NAC) prior to AngII exposure, and the protein expression of phospho-P42/44 and PDGF-B were measured with Western blotting. The DNA binding activity of EGR-1 was analyzed using electrophoretic gel mobility shift assay (EMSA), and the expression of PDGF-B was detected immunohistochemically.

RESULTSAngII induced phospho-P42/44 expression in HSC-T6, which was abrogated by U0126 or irbesartan. NAC did not inhibit phospho-P42/44 expression. EMSA showed that AngII exposure of the HSC cells markedly increased EGR-1 DNA binding activity, reaching the maximum after 60 min of exposure followed by progressive declination; irbesartan and U0126 significantly suppressed AngII-induced EGR-1 activity enhancement. ACEI at 1 micromol/L and 10 nmol/L inhibited EGR-1 activity, but ACEI at the concentration of 0.1 nmol/L resulted in enhanced EGR-1 activity. NAC showed no obvious effect in suppressing EGR-1 activity. AngII increased PDGF-B protein level in the HSCs, the effect of which was inhibited by irbesartan. U0126, NAC and ACEI did not attenuate PDGF-BB protein level in the HSCs.

CONCLUSIONStimulation of the HSCs with AngII results in EGR-1 activation via the ERK1/2 pathway, leading to up-regulation of PDGF-B expression.

Angiotensin II ; pharmacology ; Blotting, Western ; Cells, Cultured ; Early Growth Response Protein 1 ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Humans ; Immunohistochemistry ; Platelet-Derived Growth Factor ; biosynthesis ; Proto-Oncogene Proteins c-sis ; Signal Transduction ; drug effects