Imperatorin ameliorates learning and memory deficits through BDNF/TrkB and ERK/CaMKIIα/CREB signaling in prenatally-stressed female offspring.

Prenatal stress (PS) can lead to impaired spatial learning and memory in offspring. Imperatorin (IMP) is a naturally occurring furanocoumarin with many pharmacological properties. However, the effects of IMP on cognitive impairment induced by PS and the underlying molecular mechanisms remain unclear. We investigated the protective effect of IMP treatment after PS on learning and memory deficits in female offspring at postnatal 60 days. After treating prenatally-stressed offspring with IMP (15 and 30 mg/kg) for 28 days, we found that IMP increased body weight and ameliorated spatial learning and memory and working memory deficits in female offspring rats. Meanwhile, hippocampal Glu and serum corticosterone levels in prenatally-stressed offspring were significantly decreased after IMP administration. Additionally, IMP treatment significantly increased BDNF, TrkB, CaMKII, and CREB mRNA expression in the hippocampus of offspring rats. Furthermore, PS-mediated induction of RKIP protein and mRNA expression and glucocorticoid receptor protein expression in the hippocampus of offspring rats were significantly decreased by IMP treatment, and the protein expression of BDNF and TrkB and relative levels of p-EKR/ERK, p-CaMKIIα/CaMKIIα, and p-CREB/CREB were remarkably increased after IMP treatment. Taken together, IMP can ameliorate PS-induced learning and memory deficits through BDNF/TrkB and ERK/CaMKIIα/CREB signaling pathway and hypothalamic-pituitary-adrenal axis.

Tra2ß protects against the degeneration of chondrocytes by inhibiting chondrocyte apoptosis via activating the PI3K/Akt signaling pathway.

OBJECTIVE: Osteoarthritis (OA) is a common disease in the elderly and seriously affects the quality of life of patients. Tra2ß is a protein that has been found to activate PI3K/Akt in recent years. The purpose of this study was to explore the protective effects of Tra2ß on chondrocytes and its mechanisms. PATIENTS AND METHODS: The expression of Tra2ß in knee cartilage tissue of patients with OA and normal people was compared. In addition, human primary chondrocytes were cultured, the expression of Tra2ß in chondrocytes by cell transfection was changed, and its effects on extracellular matrix, inflammation, and apoptosis in chondrocytes were examined. LY294002 was also used to inhibit the activity of PI3K/Akt signaling pathway to verify the mechanism of Tra2ß to protect chondrocytes. RESULTS: The expression of Tra2ß in the cartilage tissue of the OA group was significantly lower than that of the control group, and the IL-1ß-induced chondrocytes also expressed the lower Tra2ß. The overexpression of Tra2ß increased the expression of extracellular matrix collagen II and decreased the expressions of MMP3/13, inflammatory factors (IL-6, IL-8 and TNF-α), and apoptotic factors (caspase3/9, Bax). In addition, the overexpression of Tra2ß also increased expression and phosphorylation of PI3K and Akt. However, LY294002 attenuated the protective effect of Tra2ß on chondrocytes by inhibiting the PI3K/Akt signaling pathway. CONCLUSIONS: Tra2ß activates the PI3K/Akt signaling pathway, reduces the degradation of extracellular matrix of chondrocytes, reduces the level of inflammation and apoptosis of chondrocytes, and thus, plays a role in the treatment of OA.

MRI-based investigation on outflow segment of cerebral venous system under increased ICP condition.

OBJECTIVE: Increased intracranial pressure (ICP) is responsible for causing most nervous system diseases to progress seriously, till death. Recently, volume-targeted therapeutic strategy against increased ICP, which works by releasing excessive intracranial liquid especially from the venous compartment, attracted a great deal of attention. Previous research by us found a structurally special "outflow segment cuff" that is located at the juncture of superior sagittal sinus (SSS) and the brain-bridging veins in porcine model. Sequential observation demonstrated that this special structure appeared to have functional abnormalities. Based on these findings, it was proposed to try and prove a further hypothesis that there exists a similar structure in human beings that might be of importance for cerebral venous system to intervene in volume-initiated ICP regulation. Meanwhile, the diameters of bridging veins under either increased or normal ICP are compared by means of magnetic resonance imaging (MRI). METHOD: Forty patients who presented with increased ICP were selected to undergo 2D time of flight (TOF) venography and ten normal volunteers were taken as the control group. Increased intracranial pressure status was evaluated by using flash visual evoked potential (fVEP) technique. All the patients and volunteers underwent 2D-TOF MRI imaging for the following parameters: repetition time/echo time, 50/4.9 milliseconds; flip angle, 45 degrees ; field of view, 250x250 mm; matrix, 256x256 pixels; section thickness, 1.5 mm. Syngo fastview imaging system was used to process and analyze the targeted brain-bridging venous section. RESULTS: By using 2D-TOF method in vivo, most bridging venous profiles as well as SSS and vicinal cortical veins could be clearly visualized. A short and narrow section, as previously described, obviously emerged because of MRI signal weakness even disappearing at the juncture of SSS and bridging veins in increased ICP patients. In combination with previous animal morphological findings we believe that this section with abnormal MRI signal could stand for the human counterpart of "outflow segment cuff" in porcine. Such a special structure could be observed within a majority of increased ICP patients (32/40 cases), whereas only one case presented the existence of similar imaging signal weakness. Furthermore, the diameters of the bridging veins in increased ICP group are statistically larger than the control group. CONCLUSION: Intracranial venous compartment occupies about 70 to 80% blood volume inside the inflexible cranial cavity. Following volume-targeted rationale, ICP can be regulated effectively by the fluctuation of venous blood volume based on different aspects of morphology, biomechanics, and hemodynamics. In the present study, the coincidence of animal model and human venography in vivo offers strong evidences to support the hypothesis that venous hemodynamics, although passively, influences intracranial pressure environment through a possible key regulator - outflow segment narrow structure. The fact that this narrow formation and proximal vascular dilation appears more in patients under high ICP condition rather than in patients with normal pressure. Both narrow formation and proximal vascular dilation indicate its significant contribution to intracranial venous congestion, resulting from difficult drainage and the close relationship between intracranial venous volume and ICP.

Presence of exon splicing silencers within human immunodeficiency virus type 1 tat exon 2 and tat-rev exon 3: evidence for inhibition mediated by cellular factors.

Human immunodeficiency virus type 1 (HIV-1) pre-mRNA splicing is regulated in order to maintain pools of unspliced and partially spliced viral RNAs as well as the appropriate levels of multiply spliced mRNAs during virus infection. We have previously described an element in tat exon 2 that negatively regulates splicing at the upstream tat 3' splice site 3 (B. A. Amendt, D. Hesslein, L.-J. Chang, and C. M. Stoltzfus, Mol. Cell. Biol. 14:3960-3970, 1994). In this study, we further defined the element to a 20-nucleotide (nt) region which spans the C-terminal vpr and N-terminal tat coding sequences. By analogy with exon splicing enhancer (ESE) elements, we have termed this element an exon splicing silencer (ESS). We show evidence for another negative cis-acting region within tat-rev exon 3 of HIV-1 RNA that has sequence motifs in common with a 20-nt ESS element in tat exon 2. This sequence is juxtaposed to a purine-rich ESE element to form a bipartite element regulating splicing at the upstream tat-rev 3' splice site. Inhibition of the splicing of substrates containing the ESS element in tat exon 2 occurs at an early stage of spliceosome assembly. The inhibition of splicing mediated by the ESS can be specifically abrogated by the addition of competitor RNA. Our results suggest that HIV-1 RNA splicing is regulated by cellular factors that bind to positive and negative cis elements in tat exon 2 and tat-rev exon 3.

The exon splicing silencer in human immunodeficiency virus type 1 Tat exon 3 is bipartite and acts early in spliceosome assembly.

Inefficient splicing of human immunodeficiency virus type 1 (HIV-1) RNA is necessary to preserve unspliced and singly spliced viral RNAs for transport to the cytoplasm by the Rev-dependent pathway. Signals within the HIV-1 genome that control the rate of splicing include weak 3' splice sites, exon splicing enhancers (ESE), and exon splicing silencers (ESS). We have previously shown that an ESS present within tat exon 2 (ESS2) and a suboptimal 3' splice site together act to inhibit splicing at the 3' splice site flanking tat exon 2. This occurs at an early step in spliceosome assembly. Splicing at the 3' splice site flanking tat exon 3 is regulated by a bipartite element composed of an ESE and an ESS (ESS3). Here we show that ESS3 is composed of two smaller elements (AGAUCC and UUAG) that can inhibit splicing independently. We also show that ESS3 is more active in the context of a heterologous suboptimal splice site than of an optimal 3' splice site. ESS3 inhibits splicing by blocking the formation of a functional spliceosome at an early step, since A complexes are not detected in the presence of ESS3. Competitor RNAs containing either ESS2 or ESS3 relieve inhibition of splicing of substrates containing ESS3 or ESS2. This suggests that a common cellular factor(s) may be required for the inhibition of tat mRNA splicing mediated by ESS2 and ESS3.

Pretreatment With Erythropoietin Attenuates Intestinal Ischemia Reperfusion Injury by Further Promoting PI3K/Akt Signaling Activation.

BACKGROUND: Erythropoietin (EPO) has been shown to be beneficial in resolution of acute inflammation and intestinal ischemia/reperfusion (IR) injury is featured by the excessive immune response. The current research is designed to evaluate the effect and potential mechanisms of EPO on the intestinal IR injury. Therefore, the effect of EPO on intestinal IR injury was examined by the change of intestinal histology; the expression of pro-inflammatory cytokines tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and interferon γ (IFN-γ); and the protein levels of EPOR, p-EPOR, p85, p-p85, Akt, p-Akt, IκΒ-α, p-p65, and p65. MATERIALS AND METHODS: Thirty male Sprague-Dawley rats were randomly divided into three groups: sham group (sham), IR-saline group (IRI), and the IR-EPO group (EPO). Rats were treated with EPO (5000 U/kg) 1 hour before IR induction. A rat model of IR injury was established by ligating the superior mesenteric artery for 30 minutes, followed by reperfusion for 1 hour. Intestinal histology, pro-inflammatory cytokines, and mediators were assessed. The effect of EPO on PI3K/Akt/NF-κB signaling and EPOR were also measured. RESULTS: EPO significantly decreased the pathologic changes of intestinal and reduced the elevation of pro-inflammatory cytokines TNF-α, IL-1ß, and IFN-γ in intestinal and serum caused by IR which was associated with suppressing NF-κB activation by further promoting activation of PI3K/Akt signaling. CONCLUSIONS: EPO ameliorated the acute intestinal injury caused by IR, which was associated with further activating PI3K/Akt signaling to suppress NF-κΒ-mediating inflammation. Our findings suggest that EPO could be useful for preventing IR-induced intestinal injury.

Expression and regulation of glucokinase in rat islet beta- and alpha-cells during development.

Glucokinase (GK) is the rate-limiting enzyme in the glycolytic pathway of the beta-cell and, even in the rat fetus at 22-days gestation, immediately before birth, acts as a sensor of glucose influencing the rate of glucose utilization. However, when GK first appears in islets during beta-cell development is unknown. Whether GK is expressed in fetal glucagon-producing cells is also unknown. To determine this information, fetal rat islets were examined at 16-, 18-, and 22-days gestation. GK was identified immunocytochemically in both beta- and alpha-cells at all these ages, with the number of GK immunoreactive cells positively correlated to the fetal age from 16-22 days. Western blot analysis of islet protein extracts demonstrated the presence of GK, at 52 kDa, at 16 days and thereafter. To determine whether glucose had any effect on regulation of GK biosynthesis, fetal islets were cultured in medium containing a wide range of concentrations of glucose for 7 days. The amount of GK protein was significantly decreased in low concentrations of glucose and augmented at high concentrations. In conclusion, GK was expressed in both beta- and alpha-cells in fetal rat islets during development. GK is an integral part of the function of both of these cells at all stages in the development of the fetal islet.

Single-nucleotide polymorphisms in the lipoprotein lipase gene associated with coronary heart disease in Chinese.

Coronary heart disease is a complex disease reflecting the interaction of multiple genes with the environment (e.g. diet, life style). Lipoprotein lipase (LPL) plays an important role in lipid metabolism and the pathogenesis of coronary atherosclerosis. Recent associations between single-nucleotide polymorphisms in the LPL gene and heart disease have been reported, but little is known in Chinese. The LPL gene spans >26 kb, with an mRNA of 3549 bp. In the present study, we screened 5155 bp (565 bp of 5' flanking region, nine exons and donor- and acceptor-splice sites, and some intronic bases) in 160 Chinese patients with confirmed coronary heart disease and 150 age- and gender-matched controls. Thirteen of the sixteen single-nucleotide polymorphisms that we found have not been previously reported. In males, significant (P<0.05) differences between the coronary heart disease patients and controls were found for five single-nucleotide polymorphisms: -421G>A (5' flanking region); +13,577C>A (intron 2); +16,052G>A, R192Q (exon 5); +16,173C>G and +16,177T>C (intron 5). In females, significant differences between the patients with coronary heart disease and controls were found for only the -421G>A and +16,052G>A (R192Q) mutations. Among the coronary heart disease males, significant (P<0.05) associations were found between the low-HDL high-triglyceride (LHDL/HTG) phenotype and the non-LHDL/HTG trait for the 5' flanking-421G, the intron 2+13,577C, and the exon 5+16,052G mutations, with odds-ratios (ORs)[confidence intervals] of 3.90[1.12-13.66], 3.38[1.22-9.40], and 3.22[1.04-10.01], respectively; no corresponding associations were found in females. There were 69, 51, 57 and 41 unphased haplotype patterns in male coronary heart disease, male control, female coronary heart disease and female control groups, respectively; the computer program PM-Plus found the heterogeneity model by far the best fit (P<0.0001 in males, >0.01 in females). These data show that some single-nucleotide polymorphisms in the LPL gene among Chinese are associated with abnormal lipid and lipoprotein profiles and predisposition to coronary heart disease, a genetically heterogeneous complex disease, and that they are gender-specific.

Clinical responses and lymphoid infiltrates in metastatic melanoma following treatment with intralesional GM-CSF.

Past studies in animal models with gene-transfected tumour cells have suggested that GM-CSF may have a role in immunotherapy of tumours as a result of the effects it has on antigen-presenting cells. The present (phase I) studies were carried out to examine whether intralesional injections of GM-CSF induce regression of subcutaneous metastases in patients with melanoma and influence lymphoid infiltrates in and around the metastases. Thirteen patients had 15-50 mg doses of GM-CSF injected into two subcutaneous metastases. In each case one metastasis received only five injections before excision whereas the other received weekly injections up to 6 months. Partial regression of injected and/or non-injected metastases was seen in three patients. The metastases from the responding patients that were treated with intralesional GM-CSF had marked increases and high absolute numbers of T cell infiltrates into the tumour, particularly of the CD4 T cell subset. There was an increase in IL-2R expression on the T cells and an increase in the number of Langerhans' cells infiltrating the tumours. The best predictors of clinical responses therefore appeared to be high relative increases and high absolute numbers of CD4+ T cells and Langerhans' cells within the treated tumour. These results provide support for further exploration of the role of GM-CSF in immunotherapy of human melanoma.

Immunohistological examination of the relationship between metastatic potential and expression of adhesion molecules and 'selectins' on melanoma cells.

Identification of antigens by monoclonal antibodies (MAbs) on sections of human melanoma by immunoperoxidase techniques was used to determine whether certain adhesion molecules and "selectin-like" molecules may be related to the metastatic potential of primary melanoma. The adhesion molecules examined were the leukocyte function antigen (LFA-1) and its ligand--intercellular adhesion molecule-1 (ICAM-1), the receptor alpha V beta 3 for vitronectin, its subunits alpha V and beta 3, and the CD36 receptor for thrombospondin (TSP). The criteria used to establish metastatic potential were relation of the molecules to tumor thickness and differences in expression: (i) between radial and vertical growth phases of the primary tumors and (ii) between 34 primary and 21 unrelated metastases. By these criteria ICAM-1, alpha V beta 3 and its subunit were associated with the malignant potential of primary melanoma. These molecules were not expressed on nevi or other skin cancers with low metastatic potential such as squamous (SCC) and basal cell carcinomas (BCC). In contrast, expression of TSP and the CD36 receptor for TSP were not related to metastatic potential. CD36 was expressed widely not only on melanoma but also on BCC, SCC and nevi. Similarly, the selectin-like molecule, CD44, was widely expressed on melanoma and non-melanoma carcinomas. The lymph node homing receptor, Leu 8, and the cutaneous lymphocyte antigen (CLA) were not detected on melanoma. Leu 8 was present on normal epithelium and SCCs, and common leucocyte antigen (CLA) was detected on lymphocytes in the epithelium and near melanoma. These results support previous suggestions that expression of ICAM-1 and V beta 3 integrin or its subunit beta 3 on melanoma may be a useful prognostic marker in primary melanoma. They do not support a role for CD44, Leu 8, CLA and TSP or its receptor CD36 in the metastatic process in melanoma.

Simultaneous determination of neodymium, erbium and holmium in rare earth mixtures with 2-phenyltrifluoroacetone and octylphenol poly(ethyleneglycol) ether by third-derivative spectrophotometry.

The complexes of the rare earth metals with 2-phenyltrifluoroacetone in the presence of TX-100 are reported. The characteristic absorbances of neodymium, holmium and erbium complexes can be increased by factors of 8.5, 31 and 15 respectively, compared to those of the cholrides. The third-derivative spectra have been used to eliminate the interference of cerium, and the sensitivities are increased again by factors of 7.4, 5.5 and 6.5. A method for the direct determination of neodymium, erbium and holmium in rare earth mixtures is proposed.

Collagen I/III and matrix metalloproteinases (MMP) 1 and 13 in the fascia of patients with incisional hernias.

The late appearance ofincisional hernias several years after laparotomy and the high recurrence rates after operation strongly imply the presence of a disorder of the connective tissue, although a specific defect in patients with incisional hernias has not yet been identified. In the present study we used both immunohistochemistry and Western blot analysis to evaluate the ratio of collagen I and III and the expression of the metalloproteinases (MMP) 1 and 13 in the fascia of patients with incisional or recurrent incisional hernias. Samples of healthy skin or stable skin scar in patients without hernias served as controls. Altogether, our data indicated a significantly decreased ratio of collagen I/III in the fascia of patients with incisional hernias and recurrent incisional hernias. Furthermore, in these patients the expression of MMP-1 was decreased compared to the controls, whereas MMP-13 could not be detected in any fascia sample, with or without hernias present. For the first time, our results give evidence of the existence of a possible collagen disorder in these patients. The decreased ratio ofcollagen I/III is explainable due to a relative increase of collagen type III, which is known to be characterized by thin fibril diameters and lowered mechanical strength. The altered collagen ratio might be the result of the decreased activity of MMP-1, whereas the absent MMP-13 expression did not seem to modify the scar formation. Thus, our data indicate the presence of collagen metabolic disorders in patients with incisional hernias and recurrent incisional hernias. Furthermore, these results might explain the poor results of a mesh-free hernia repair, which again builds up scar tissue of inadequate collagen composition and strength.

Absorption spectra of the 4f electron transitions of the praseodymium complex with 1-cyclopropyl-6-fluoro-1,4-dihydro-7-(4-ethyl-1-piperazinyl) -4-oxo-3-quinoline carboxylic acid hydrochloride and its analytical application.

The absorption spectra of the praseodymium complex with 1-cyclopropyl-6-fluoro-1,4-dihydro-7-(4-ethyl-1-piperazinyl)-4-oxo-3- quinoline carboxylic acid hydrochloride (NNFX) has been studied by normal and derivative spectrophotometry. The complex showed maximum absorption at 350 nm at pH 6.0. The stoichiometry of the Pr-NNFX complex was calculated by the molar ratio and continuous variation methods. The ratio of Pr to NNFX was 1:3. The absorption bands of the 4f electron transitions of the praseodymium complex with NNFX are enhanced markedly, especially the wavelength at 481 nm. Using the third-derivative spectrum, the calibration graph is linear over the range 2.5 x 10(-5)-3.5 x 10(-4) mol dm-3 for praseodymium. The detection limits (signal-to-noise ratio of 2) is 1.4 x 10(-6) mol dm-3. The relative standard deviation is 1.2% for 7.0 x 10(-5) mol dm-3 of praseodymium. A method for the direct determination of praseodymium in rare earth mixtures with good accuracy and selectivity is described.

[Fluorescence enhancement of the dysprosium-sulfosalicylic acid system by lanthanum and its application].

A fluorescence enhancement produced by adding lanthanum(III) to dysprosium-sulfosalicylic acid system was observed. La3+ enhanced the fluorescence intensity by two orders of magnitude compared with the system without La3+. The system was used for the determination of trace amount of dysprosium in the range of 3.0 x 10(-7)-1.0 x 10(-5) mol/dm3, and the detection limit was 8.0 x 10(-8) mol/dm3. The procedure was applied to the determination of dysprosium in synthetic rare earth samples and standard rare earth samples with satisfactory results.

[Studies on morphology and histology of medicinal part of Corydalis yanhusuo].

OBJECTIVE: To Study the morphology, microstructure and morphosis of medicinal part of Corydalis yanhusuo, and provide a basis for the knowledge of the type of modified stem of Corydalis yanhusuo and the identification of crude drug. METHOD: Sample collection, culture observation and microsectile observation. RESULT: Morphological and histological characters and morphogenetic and developmental regulation of medicinal part of Corydalis yanhusuo are described with pictures. CONCLUSION: The medicinal part of Corydalis yanhusuo is bulb, and the conception that the medicinal part of Corydalis yanhusuo is tuber is wrong.

Reproducibility of anterior chamber angle measurement using the Tongren ultrasound biomicroscopy analysis system.

PURPOSE: To assess the intraobserver and interobserver variability of new software for the analysis of anterior chamber-angle images obtained using ultrasound biomicroscopy (UBM). PATIENTS AND METHODS: Consecutive UBM images of 4 quadrants (the superior, the nasal, the temporal, and the inferior) of 30 primary angle-closure suspects (PACSs) and 30 normal subjects with open angles were selected. Two observers used the new UBM image analysis software to calculate the following parameters on 2 separate occasions: angle opening distance (AOD), trabecular-iris angle (TIA), angle recess area (ARA), trabecular-ciliary process distance (TCPD), iris-ciliary process distance (ICPD), and iris thickness (IT). Intraobserver and interobserver reproducibility were measured using intraclass correlation coefficient (ICC) and Bland-Altman plots. RESULTS: For intraobserver measurements in PACS, the range of ICC for AODs, TIAs, ARAs, TCPDs, ICPDs, and ITs in the 4 quadrants were 0.77 to 0.94, 0.80 to 0.92, 0.82 to 0.94, 0.51 to 0.73, 0.42 to 0.78, and 0.76 to 0.89; in open angles, these were 0.83 to 0.94, 0.80 to 0.93, 0.84 to 0.93, 0.57 to 0.89, 0.57 to 0.84, and 0.70 to 0.95. For interobserver measurements in PACS, the range of ICC for AODs, TIAs, ARAs, TCPDs, ICPDs, and ITs in the same quadrants as above were 0.52 to 0.89, 0.65 to 0.92, 0.68 to 0.92, 0.22 to 0.80, 0.48 to 0.93, and 0.29 to 0.75; in open angles, these were 0.57 to 0.87, 0.60 to 0.86, 0.64 to 0.90, 0.27 to 0.94, 0.56 to 0.93, and 0.28 to 0.66. CONCLUSIONS: Angle width (AOD, TIA, and ARA) and IT of both PACS and open angles could be reliably measured by the same observer; interobserver measurements and those related to the ciliary process (TCPD and ICPD) were more variable. For the current version of this software, we recommend measurements of anterior chamber parameters by the same observer.

Gastroduodenal arterial reconstruction of the pancreaticoduodenal allograft.

Simultaneous procurement of the pancreas and liver necessitates division of vessels supplying both organs. The integrity of the pancreatic arterial supply appears to be related to surgical complications after pancreas transplantation. We have described herein three cases of gastroduodenal artery (GDA) reconstruction during pancreas transplantation, and reviewed other options for GDA reconstruction. These techniques performed safely during bench reconstruction can be applied to various clinical situations.