Contrast-induced encephalopathy with significantly elevated levels of cerebrospinal fluid protein.

Contrast-induced encephalopathy (CIE) is a rare complication of angiography. According to our knowledge, the majority of CIE reports is imaging observations and rarely includes results of cerebrospinal fluid (CSF) tests. Furthermore, among the cases reporting the data for CSF testing, most of the results were normal. Here, we report a case of CIE presenting with significantly elevated levels of CSF protein. We found that the course of improvement in brain imaging findings was not consistent with the severity of clinical manifestations. The diffusion-weighted imaging (DWI) and apparent diffusion coefficient (ADC) sequences were normal. Considering the lack of convenient direct indicators to observe blood-brain barrier (BBB) function, changes in the levels of CSF protein may be related to BBB permeability and recovery and may serve as a potential prognostic marker.

Genomic insights into mite phylogeny, fitness, development, and reproduction.

BACKGROUND: Predatory mites (Acari: Phytoseiidae) are the most important beneficial arthropods used in augmentative biological pest control of protected crops around the world. However, the genomes of mites are far less well understood than those of insects and the evolutionary relationships among mite and other chelicerate orders are contested, with the enigmatic origin of mites at one of the centres in discussion of the evolution of Arachnida. RESULTS: We here report the 173 Mb nuclear genome (from 51.75 Gb pairs of Illumina reads) of the predatory mite, Neoseiulus cucumeris, a biocontrol agent against pests such as mites and thrips worldwide. We identified nearly 20.6 Mb (~ 11.93% of this genome) of repetitive sequences and annotated 18,735 protein-coding genes (a typical gene 2888 bp in size); the total length of protein-coding genes was about 50.55 Mb (29.2% of this assembly). About 37% (6981) of the genes are unique to N. cucumeris based on comparison with other arachnid genomes. Our phylogenomic analysis supported the monophyly of Acari, therefore rejecting the biphyletic origin of mites advocated by other studies based on limited gene fragments or few taxa in recent years. Our transcriptomic analyses of different life stages of N. cucumeris provide new insights into genes involved in its development. Putative genes involved in vitellogenesis, regulation of oviposition, sex determination, development of legs, signal perception, detoxification and stress-resistance, and innate immune systems are identified. CONCLUSIONS: Our genomics and developmental transcriptomics analyses of N. cucumeris provide invaluable resources for further research on the development, reproduction, and fitness of this economically important mite in particular and Arachnida in general.

A sensitive, non-damaging electrochemiluminescent aptasensor via a low potential approach at DNA-modified gold electrodes.

Electrochemiluminescence (ECL)-based biosensors are often used in the field of DNA- and protein-assay. Although ruthenium complex-based ECL is sensitive, its high exciting potential may lead to oxidation damage to biomolecules. For the first time, a non-damaging, low potential ECL aptasensor was constructed for bioassay with lysozyme as a model. After a single-stranded anti-lysozyme aptamer was attached to a gold electrode, a double stranded (ds)-DNA formed with its complementary strand. Ru(phen)(3)(2+), as an ECL probe, was intercalated into the ds-DNA. The hybridization of lysozyme with its aptamer led to the dissociation of ds-DNA because of the high stability of the aptamer-lysozyme and therefore the Ru(phen)(3)(2+) intercalated into ds-DNA was released. A low potential ECL was observed at the ds-DNA-modified electrode because ds-DNA was able to preconcentrate tripropylamine (TPA) and acted as the acceptor of the protons released from protonated TPAH(+). While the DNA sequence (anti-lysozyme aptamer) was used as the special recognition element for lysozyme, the formed ds-DNA also provided a micro-environment for low potential ECL. The low potential ECL aptasensor achieved the determination of lysozyme with a detection limit of 0.45 pM. The day-to-day precision (RSDs, n = 5) for the determination of lysozyme was lower than 5%, showing the reliability of the aptasensor. The regeneration of the aptasensor confirmed that the low potential for ECL could decrease oxidation damage to biomolecules. Further, the proposed method was successfully used to analyze diluted egg white sample directly. The protocol exhibited a promising platform for sensitive bioassay and could be further applied for the development of other low potential ECL sensing systems.

The electrochemiluminescence of ruthenium complex/tripropylamine systems at DNA-modified gold electrodes.

The electrochemiluminescence (ECL) behavior of ruthenium complex/tripropylamine (TPA) systems at DNA-modified gold electrode was studied to understand the possible mechanism and to develop new detection platforms. DNA strand, especially double-stranded DNA (ds-DNA), can preconcentrate TPA and acts as the acceptor of the protons released from TPAH(+), therefore the improved ECL emission and the low potential ECL were observed. The intercalation of Ru(phen)(3)(2+) into ds-DNA was confirmed to be a sensitive and label-free DNA-related detection platform. The above results were validated by the analysis of lysozyme using anti-lysozyme aptamer-modified electrode. This work opens a new field by the use of DNA-modified electrode to develop novel sensing platforms, such as low potential ECL biosensors and Ru(phen)(3)(2+) intercalation-based ECL biosensors.

Electrochemical aptasensor using the tripropylamine oxidation to probe intramolecular displacement between target and complementary nucleotide for protein array.

Tripropylamine (TPA) has different oxidation efficiency at double stranded (ds)-and single stranded (ss)-DNA-modified electrodes. Using this property, a simple but sensitive biosensor using TPA oxidation to probe the intramolecular displacement was constructed with the analysis of lysozyme as model for the first time. After the complementary ss-DNA strand of anti-lysozyme aptamer was immobilized onto gold electrode via gold-thiol bond, the incubation with the aptamer resulted in the formation of ds-DNA. Lysozyme (in 10 µL sample) binding with aptamer displaced the complementary strand because of the high affinity of lysozyme and its aptamer, corresponding to the dissociation of the ds-DNA. The modified electrode was swept in 20mM TPA solution from 0.2 to 0.95 V. The difference in oxidation current was used to quantify the content of lysozyme with a linear range from 1.0 pM to 1.1 nM. That means 10 amol or 6.0 × 10(6) lysozyme molecules can be detected. Because the signal is produced from the preconcentrated TPA at the electrode surface, the high sensitivity is achieved over the single site labelling strategy. The proposed method is simple, stable, specific, and time-saving while the complicated sample pre-treatment and the labelling to the DNA strand are avoided. The biosensor was validated by the analysis of the diluted egg white sample directly. The recovery and reproducibility were 93.3-100% and 1.4-4.2%, respectively.

Polydopamine-based permanent coating capillary electrochromatography for auxin determination.

A novel, simple, and economical method for the preparation of open-tubular capillary column using polydopamine coating was reported for the first time. After the capillary was filled with dopamine solution for 20h, polydopamine was formed and deposited on the inner wall of capillary as permanent coating via the oxidation of dopamine by the oxygen dissolved in the solution. Moreover, the electroosmotic flow of the coated capillaries was measured to be dependent on the repetitive coating times. The performance of the polydopamine-coated capillary electrochromatography was validated by the analysis of four auxins, indole-3-butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (dCPAA), indole-3-acetic acid (IAA), and phenoxyacetic acid (PAA). The precisions (RSD, n=5) were in the range of 1.6-2.4% for migration time, 4.0-6.5% for peak area response, and 3.6-4.7% for peak height response for the four auxins at 1microgmL(-1) level. The detection limits were 0.185, 0.172, 0.177, and 0.259microg/mL for IBA, dCPAA, IAA, and PAA, respectively. The method was successfully used to the determination of IAA in the culture media of IAA-producing bacteria.

[Increased levels of prostaglandin E2 and bcl-2 in peritoneal fluid and serum of patients with endometriosis].

OBJECTIVE: To detect the concentrations of prostaglandin E2 (PGE2) and bcl-2 in sera and peritoneal fluid of women with endometriosis. METHODS: The study group included 36 samples of peritoneal fluid and serum respectively from patients with endometriosis, and control group included 30 samples of peritoneal fluid and serum respectively from patients without endometriosis (either ovary cyst or uterine myoma). The peritoneal fluids were collected at the time of laparoscopic operation, and the sera were collected before surgery. Concentrations of PGE2 and bcl-2 were determined by enzyme linked immunosorbent assay (ELISA). RESULTS: The peritoneal fluid concentrations of PGE2 and bcl-2 in study group were significantly higher than that of control group, (1987 +/- 532) ng/L vs (386 +/- 215) ng/L, (177 +/- 53) U/L vs (86 +/- 21) U/L, (P < 0.05); and the PGE2 levels of severe endometriosis were significantly higher than that of mild endometriosis, (2221 +/- 1352) ng/L vs (1694 +/- 381) ng/L, (P < 0.01). The serum concentrations of PGE2 and bcl-2 in study group were significantly higher than that of control group, (3787 +/- 514) ng/L vs (129 +/- 97) ng/L, (96 +/- 44) U/L vs (53 +/- 40) U/L, (P < 0.01). Serum PGE2 concentrations of severe endometriosis were significantly higher than that of mild endometriosis, (964 +/- 290) ng/L vs (590 +/- 362) ng/L, (P < 0.01). CONCLUSIONS: The concentrations of PGE2 and bcl-2 in peritoneal fluid are increased in endometriosis. The concentrations of PGE2 and bcl-2 are associated with the extent of endometriosis lesions.

[Protective effect of follicle stimulating hormone on apoptosis of human epithelial ovarian cancer cell induced by cisplatin].

OBJECTIVE: To observe the protective effects of follicle stimulating hormone (FSH) on human epithelial ovarian cancer cell apoptosis induced by cisplatin (DDP). METHODS: OVCAR3-FSHR cell were treated with DDP and FSH at serials of concentrations, MTT assay was used to examine the growth inhibition of OVCAR3-FSHR cell after treatment with DDP and FSH. Flow cytometry was used to analyze the change of cell cycle and percentage of apoptosis. RESULTS: It was revealed that FSH decreased the growth inhibition induced by DDP. We also demonstrated that FSH reduced the S-phase percentage compared with the DDP only groups after treatment for 24 hours and reduced apoptosis percentage after 48 hours treatment with DDP. CONCLUSION: It is suggested that FSH can protect the apoptosis induced by DDP. It also suggests that FSH may be an important chemoresistent reason for the chemotherapy of ovarian cancer.

[Follicle stimulating hormone inhibits cisplatin induced apoptosis in ovarian cancer cells].

OBJECTIVE: To investigate whether the follicle stimulating hormone (FSH) can inhibit apoptosis in ovarian cancer cells induced by cisplatin (DDP) and its possible mechinism. METHODS: DNA fragmentation assay, (TdT-mediated dUTP nick end labling TUNEL), Western blot were used to analyze the changes in expression levels of Survivin and bcl-2 protein. The relative activity of caspase-3 was also determined. RESULTS: 200 mIU/ml FSH could regulate down the percentage of apoptotic cells and DNA fragmentation induced by 5.0 micrograms/ml cisplatin, while 200 mIU/ml FSH increased Survivin protein expression but could't influence the expression of bcl-2 protein. CONCLUSION: FSH can inhibit ovarian cancer cells apoptosis induced by cisplatin. The possible mechinism is up-regulation of Survivin expression and down-regulation of caspase activity.

[Preliminary study on pathway of follicle-stimulating hormone on human epithelial ovarian cancer cell proliferation].

OBJECTIVE: There is only a limited direct indication that gonadotropins play a role in the genesis and development of epithelial ovarian cancer (EOC). Follicle-stimulating hormone (FSH) can enhance the growth of epithelial ovarian cancer cell in vitro. The research is to investigate the pathway of FSH action in epithelial ovarian cancer cell. METHODS: Epithelial ovarian cancer cell line OVCAR3 was transfected with FSH receptor cDNA expressing vector. The transfected cells that are sensitive highly to FSH stimulation were got, and named OVCAR3-FSHR. Adding FSH to the cells, or treating the cells with protein kinase C (PKC) activator tetradenocanoyl phorbol acetate (TPA), PKC inhibitor tamoxifen (TAM) in meantime, methyl thiazolyl tetrazolium (MTT) method was used to study the proliferation of cells. RT-polymerase chain reaction was used to identity the mRNA expression of various PKC subtypes. Westernblot was for detection of protein expression of PKCalpha and phosphorylated PKCalpha. RESULTS: FSH can promote proliferation of OVCAR3-FSHR (1.9 folds). There is some increase in PKCalpha by the FSH stimulation. The phosphorylated PKCalpha expression were enhanced significantly too. Both the amount and activity of PKCalpha were increased in response to FSH. TPA and TAM suppress FSH-stimulated cell growth (60% and 47%). Meanwhile expression level of PKCalpha was decreased with the co-treatment of TPA or TAM and FSH comparing with treatment with FSH only. CONCLUSIONS: FSH promoted epithelial ovarian cancer cell proliferation through PKC pathway. It plays a role in the development of epithelial ovarian cancer.

Expression of vascular endothelial growth factor and thrombospondin-1 mRNA in patients with endometriosis.

OBJECTIVE: To investigate the expression of vascular endothelial growth factor (VEGF) mRNA and thrombospondin-1 (TSP-1) mRNA in endometriosis. DESIGN: Molecular studies in human tissue. SETTING: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, P. R. China. PATIENT(S): Patients undergoing laparoscopy for infertility or other benign gynecologic conditions. INTERVENTION(S): Biopsies were taken from endometriotic lesions (red peritoneal lesion, ovarian endometrioma, and unterosacral ligament nudule) and eutopic endometrium during laparoscopy. MAIN OUTCOME MEASURE(S): mRNA expression from endometriotic lesion and eutopic endometrium was analyzed by reverse transcriptase polymerase chain reaction (PCR) and Northern blotting. RESULT(S): Among the endometriotic lesions, red peritoneal lesions expressed higher levels of VEGF mRNA and lower levels of TSP-1 mRNA, whereas ovarian endometrioma expressed lower levels of VEGF mRNA and higher levels of TSP-1 mRNA. Eutopic endometrium of women with endometriosis had higher expression levels of VEGF mRNA and lower expression levels of TSP-1 mRNA than that of women without endometriosis. CONCLUSION(S): The expression of VEGF and TSP-1 in endometriotic lesions appears to be associated with the extent of their neovascularization. The imbalance in expression of VEGF and TSP-1 in the endometrium may play a role in the development of endometriosis.

Yeast one-hybrid system used to identify the binding proteins for rat glutathione S-transferase P enhancer I.

OBJECTIVE: To detect the trans-factors specifically binding to the strong enhancer element (GPEI) in the upstream of rat glutathione S-transferase P (GST-P) gene. METHODS: Yeast one-hybrid system was used to screen rat lung MATCHMAKER cDNA library to identify potential trans-factors that can interact with core sequence of GPEI(cGPEI). Electrophoresis mobility shift assay (EMSA) was used to analyze the binding of transfactors to cGPEI. RESULTS: cDNA fragments coding for the C-terminal part of the transcription factor c-Jun and rat adenine nucleotide translocator (ANT) were isolated. The binding of c-Jun and ANT to GPEI core sequence were confirmed. CONCLUSIONS: Rat c-jun transcriptional factor and ANT may interact with cGPEI. They could play an important role in the induced expression of GST-P gene.