Preeclampsia is a leading cause of maternal and infant morbidity and mortality. It is very important to explore the biomarkers for the early prediction of preeclampsia. Some peptides released from placenta, such as soluble Flt-1 and placenta growth factor (PlGF), have been revealed for definite prospects of application. Meanwhile, the recent advances in proteomics, metabolomics and microRNA shed light on searching of new biomarkers for preeclampsia prediction.

Animals ; Endothelium, Vascular ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Therapy ; Neoplasms ; blood supply ; therapy ; Neovascularization, Pathologic ; Plasmids ; Receptors, Cell Surface ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Swine

Objective To investigate the importance of plasmid-mediated quinolone resistance in the development of quinolone resistance in clinical isolates of gram-negative bacteria.Methods A total of 541 consecutive clinical isolates of gram-negative ba- cilli resistant or intermediate to ciprofloxacin were screened for the qnrA gene by PCR.Conjugation experiments were carried out with azide-resistant E.coli J53 as a recipient.The aac(6')-Ib-cr gene was detected.The mutations in the quinolone-resist- ance-determining region (QRDR) of the gyrA and parC genes were identified in qnrA positive strains.Results qnrA was identi- fied in 7 of the 541 strains.Among the qnrA positive strains,5 were Enterobacter cloacae.No qnrA was detected in nonfer- menters.Quinolone resistance was transferred in 4 of 7 qnrA positive strains.Transconjugants had 12-to 125-fold increases in MIC of ciprofloxacin relative to that of the recipient.Seven strains contained qnrA with a nucleotide sequence identical to that originally reported.Two transconjugants with higher ciprofloxacin MICs contained aac(6')-Ib-cr gene.Mutations occurred in the QRDR of the gyrA and parC genes in 5 PCR-positive clinical strains.Conclusions Transferable plasmid-mediated quinolone resistance associated with qnrA is highly prevalent in clinical strains of Enterobacter spp.aac(6')-Ib-cr gene and mutations in the quinolone targets may co-exist with qnrA,which may contribute to the further increase of resistance to quinolones.

To study the expression of HCV non-structure 5 antigen in vitro, a human HepG2 cell line was incubated with a HCV RNA positive serum. The S ABC i mmunological techniques and gold-labeled colloid electron microscopy method wer e employed to examine for the viral proteins in those cells. The HCV non-struct ure 5 antigen was first detected in the HepG2 cells at 72 hours post incubation. The antigen was continuously observed in the cytoplasm or on the membrane as we ll on the cell wall of the HepG2 cells even after 1, 2, 3 and 4 weeks post incub ation. The observation of HCV non-structure 5 antigen continuously expressed in the HepG2 cells strongly indicates that the cells may have been infected by HCV virus and the virus may have replicated in the cells. Therefore, the HepG2 cell line may be served as a potential host for establishment of HCV infection and p ropagation in vitro.

Objective To evaluate the diagnostic value of brain SPECT imaging with a novel Aβ plaque probe,131 I-2-(4'-dimethylaminophenyl) -6-iodoimidazo[ 1,2-α ] pyridine ( 131 I-IMPY) in early AD.Methods Thirteen patients with AD (3 males,10 females,age ranged 52 - 79 y),11 with mild cognitive impairment (MCI,4 males,7 females,age ranged 48 - 67 y) and 14 normal controls (6 males,8 females,age ranged 42 - 67 y) were enrolled in this study.131I-IMPY SPECT imaging was acquired in 2 -3 h after the agent injection.ROIs were drawn on cerebral lobes and cerebellum.The ratios of mean radioactivity of cerebral lobes over cerebellum (Rcl/cb) were calculated.The t-test was used for data analysis.Results In patients with MCI,Rcl/cb ratios were increased in parietal gyrus,temporal gyrus and frontal gyrus (right:1.15±0.18,1.18±0.12,1.14±0.14; left:1.16±0.11,1.19±0.18,1.15±0.09)compared with those in normal control group ( right:1.02 ± 0.12,1.05 ± 0.14,1.01 ± 0.12 ; left:1.03 ±0.13,1.05 ±0.13,1.01 ±0.14; t:2.1642 to 2.8757,all P ＜0.05).Rcl/cb ratios of basel ganglia and occipital gyms in MCI group (right:0.92 ±0.18,1.12 ±0.15; left:0.94 ±0.15,1.13 ±0.17) showed no statistical difference compared with those in normal control group (right:0.82 ±0.15,1.06 ±0.18;left:0.85 ±0.16,1.08 ±0.15; t:0.7805 to 1.4344,all P＞0.05).In patients with AD,Rcl/cb ratios were increased in parietal,temporal,basal ganglia and occipital lobes (right:1.16 ±0.19,1.24 ±0.17,1.16 ±0.13,1.14±0.11,1.23±0.10; left:1.17±0.21,1.25±0.15,1.18±0.08,1.17±0.16,1.25±0.11)compared with those in normal control group( t:2.1001 to 6.2789,all P ＜0.05).Rcl/cb ratios of parietal,temporal and frontal lobes in AD group showed no statistical difference compared with those in MCI group (t:0.1316 to 0.9806,all P ＞ 0.05 ),while Rcl/cb ratios of basal ganglia and occipital lobes in AD group were increased compared with those in MCI group ( t:2.0850 to 3.6772,all P ＜ 0.05 ).Conclusion 131 I-IMPY as a β- amyloid plaque probe for brain SPECT imaging may be potentially helpful for early diagnosis of AD.

To investigate the function of ALK3 gene, the gene regulation and the signaling pathway related to ventricular septum defect during heart development. The model mice with ALK3 gene knock-out via alpha-MHC-Cre/lox P system were bred. The mRNA expression level of control group was compared with that of experiment group and ALK3 downstream genes were screened using PCR-select cDNA subtraction microarray. The mRNA of control group was extracted from E11.5 normal mouse hearts, and that of experiment group, from E11.5 hearts of mice with alpha-MHC Cre(+/-) ALK3(F/+) genotype. It was found that the mice with ALK3 gene knock-out produced heart defects involving the interventricular septum. The platelet-activating factors acetylhydrolase and the transcription factor Pax-8 and so on, were down-regulated. However, the Protein Tyrosine Kinase (PTK) of Focal Adhesion Kinase (FAK) subfamily and beta subtype protein 14-3-3 were up-regulated in the alpha-MHC Cre(+/-) ALK3(F/-) mice. These data provide support that ALK3 gene played an important role during heart development. The platelet-activating factors acetylhydrolase and Pax-8 genes could be important ALK3 downstream genes in the BMP signaling pathway during interventricular septum development. PTK and beta subtype protein 14-3-3 might be regulatory factors in this pathway.
1-Alkyl-2-acetylglycerophosphocholine Esterase ; genetics ; metabolism ; 14-3-3 Proteins ; genetics ; metabolism ; Animals ; Bone Morphogenetic Protein Receptors, Type I ; genetics ; metabolism ; Genotype ; Heart Septal Defects, Ventricular ; genetics ; Mice ; Mice, Knockout ; Oligonucleotide Array Sequence Analysis ; PAX8 Transcription Factor ; Paired Box Transcription Factors ; genetics ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; genetics ; physiology

OBJECTIVETo investigate the effects of microRNA-144 (miR-144) expression on H9C2 (2-1) myocytes.

METHODSMiR-144 was up-regulated in primary cultured H9C2 (2-1) myocytes through transfection. Cells transfected with Lipofectamine(TM) 2000 and its mixture with miRNA synthesized randomly as blank control and negative control respectively. The up-regulation of miR-144 was confirmed by real-time PCR. Cell apoptosis was evaluated by means of CCK-8, Caspase-3 and flow cytometry.

RESULTSReal-time PCR results showed that the miR-144 expression was obviously increased in miR-144 up-regulation group (2178.84 ± 838.52) compared with negative (2.06 ± 0.73) and blank (1.00 ± 0.00) control group (all P < 0.01). The proliferation was lower, the activity of Caspase-3 was elevated and the apoptosis rates were significantly increased in miR-144 up-regulation group compared with negative and blank control group, while no significant difference was found between the latter 2 groups.

CONCLUSIONMiR-144 mimics may selectively up-regulate the expression of miR-144 in myocardial cells and consequently promote apoptosis and inhibit proliferation in myocardial cells.

Animals ; Apoptosis ; genetics ; Caspase 3 ; metabolism ; Cell Line ; MicroRNAs ; genetics ; metabolism ; Muscle Cells ; metabolism ; Rats ; Sincalide ; metabolism ; Transfection

OBJECTIVETo observe the effect of orexin-A on the recovery and cognitive function of aged rats after ketamine anesthesia.

METHODSFifty-five aged rats were divided randomly into control group, model control group, 1 nmol/L Orexin-A group, and 4 nmol/L Orexin-A group. In the latter 3 groups, the rats received an intraperitoneal injection of ketamine at 100 mg/kg, and normal saline was injected in the control group. Ten minutes after the injections, the rats received intraventricular injections of artificial cerebrospinal fluid (control and model control group) or of 10 microl 1 or 4 nmol/L Orexin-A as indicated. The behavioral changes of the rats were assessed by the duration of loss of righting reflex (LORR). Electroencephalogram (EEG) recordings were used to evaluate the changes in rat brain activity by comparison of the percent of sigma wave in EEG before and after the intraventricular injections. Morris water maze was used to test the learning and spatial localization abilities of the rats.

RESULTSKetamine resulted in obvious impairment of learning and memory abilities of the aged rats. Orexin-A at 4 nmol/L induced significant decrease in the duration of LORR and marked reduction of sigma activities in anesthetic rats (P<0.05), and obviously improved the learning and spatial localization abilities of the rats after anesthesia (P<0.05).

CONCLUSIONOrexin-A can promote the recovery and improve the cognitive function of aged rats after ketamine anesthesia.

Aging ; Anesthesia Recovery Period ; Anesthetics, Dissociative ; Animals ; Cognition ; drug effects ; Delayed Emergence from Anesthesia ; prevention & control ; Intracellular Signaling Peptides and Proteins ; pharmacology ; Ketamine ; Male ; Neuropeptides ; pharmacology ; Orexins ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo determine the affinity of new opioid receptor ligands to cloned mu opioid receptors stably expressed in CHO cell.

METHODSThe binding characteristics of the opioid ligand [3H] diprenorphine (3H-dip) were studied by cellular biological techniques and radioligands binding in cloned mu opioid receptors stably expressed in CHO cells in saturation binding experiments, and were followed by competition binding experiments with a variety of new synthesized opioid receptor ligands.

RESULTSThe Kd and Bmax of [3H] diprenorphine bound to mu receptors were 1.06 nmol/L and 930 fmol/mg protein, respectively. Competition binding experiments revealed that ligand 3# and 12# displayed much higher affinity than DAMGO and Morphine for the cloned mu opioid receptor. However, the affinities of ligands 2#, 6#, 8# and 9# were lower than DAMGO and Morphine.

CONCLUSIONThe present results suggest that the new ligands 3# and 12# have higher affinity to mu opioid receptors. However, ligands 2#, 6#, 8# and 9# have lower affinity to mu opioid receptors.

Animals ; Binding Sites ; Binding, Competitive ; CHO Cells ; metabolism ; Cloning, Molecular ; Cricetinae ; Diprenorphine ; pharmacology ; Ligands ; Receptors, Opioid, mu ; biosynthesis ; genetics ; metabolism

OBJECTIVETo observe the effects of paired box gene 8 (Pax-8) silencing by RNA interference on mitochondrial function and cardiomyocytes apoptosis.

METHODSThe cultured H9C2 (2-1) myocytes were divided into 3 groups: short interference RNA targeting Pax-8 (Pax-8 siRNA) group, non-specific siRNA group as the negative control (NC siRNA), and blank control group (BC siRNA). Fluorescence spectrophotometry was used to detect the activity of caspase-3. RT-PCR was performed to detect mRNA expression of Bcl2 and Bax. The protein expression of Bcl2, Bax and cytoplasm of Cytochrome was examined by Western blot. Changes of ΔΨm were detected by flow cytometry.ΔΨm with JC-1 monomer/polymer ratio was calculated for measuring mitochondrial depolarization proportion.

RESULTSCompared to NC siRNA and BC siRNA group (0.075 ± 0.021, 0.072 ± 0.019), the activity of caspase-3 in Pax-8 siRNA group (0.167 ± 0.012) was significantly increased (P < 0.05); Bcl2 mRNA and protein expression in Pax-8 siRNA group (0.61 ± 0.06, 0.94 ± 0.11) were significantly downregulated compared with NC siRNA group (0.90 ± 0.070, 1.39 ± 0.15) and BC siRNA group (0.94 ± 0.087, 1.49 ± 0.20) (P < 0.05); Bax mRNA and protein expression in Pax-8 siRNA group (1.05 ± 0.10, 1.25 ± 0.12) were markedly upregulated compared with NC siRNA group (0.72 ± 0.03, 0.99 ± 0.12) and BC siRNA group (0.64 ± 0.03, 0.92 ± 0.06), P < 0.05; cytosolic cytochrome expression in Pax-8 siRNA group (0.75 ± 0.14) was significantly upregulated compared with NC siRNA group (0.51 ± 0.06) and BC siRNA group (0.48 ± 0.07) (P < 0.05); JC-1 monomer/polymer ratio in Pax-8 siRNA group (0.163 ± 0.011) was significantly increased compared with NC siRNA group (0.092 ± 0.015) and BC siRNA group (0.072 ± 0.025) (P < 0.05) indicating mitochondrial membrane potential was significantly reduced in Pax-8 siRNA group. Above parameters were similar between NC siRNA group and BC siRNA group (P > 0.05).

CONCLUSIONInhibiting Pax-8 results in enhanced cardiomyocytes apoptosis through the mitochondrial pathway.

Animals ; Apoptosis ; Cells, Cultured ; Mitochondria, Heart ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; PAX8 Transcription Factor ; Paired Box Transcription Factors ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; Rats ; Transfection