Childhood maltreatment on young adult depression: A moderated mediation model of negative automatic thoughts and self-compassion.

This study examines the mediating role of negative automatic thoughts on the link between childhood maltreatment and young adult depression, and the moderating role of self-compassion in this indirect link. College students (N = 578) completed self-report questionnaires assessing the mentioned study variables. The results showed that childhood maltreatment was positively associated with young adult depression via negative automatic thoughts. Moreover, self-compassion moderated this indirect link such that participants with low self-compassion demonstrated a stronger indirect link than those with high self-compassion. These findings highlight the important role of self-compassion in countering the adverse outcomes of childhood maltreatment.

Biotransformation of ursolic acid by Alternaria longipes AS3.2875.

Microbial transformation of ursolic acid (1) was carried out by Alternaria longipes AS 3.2875. Six transformed products (2-7) from 1 were isolated and their structures were identified as 3-carbonyl ursolic acid 28-O-ß-D-glucopyranosyl ester (2), ursolic acid 3-O-ß-D-glucopyranoside (3), ursolic acid 28-O-ß-D-glucopyranosyl ester (4), 2α, 3ß-dihydroxy ursolic acid 28-O-ß-D-glucopyranosyl ester (5), 3ß, 21ß dihydroxy ursolic acid 28-O-ß-D-glucopyranosyl ester (6), and 3-O-(ß-D-glucopyranosyl)- ursolic acid 28-O-(ß-D-glucopyranosyl) ester (7) based on the analysis of 1D NMR, 2DNMR and MS data. The product 2 was a new compound among them and showed stronger antibacterial activity against S. aureu, MRSA and MRCA than substrate. In this study, we modified structure of ursolic acid through biotransformation to enhance its activities and preliminarily discussed the transformation way of the products.

HuR posttranscriptionally regulates early growth response-1 (Egr-1) expression at the early stage of T cell activation.

T cell activation depends on appropriate and precise regulation of gene expression. Here we find that rapidly translocated RNA-binding protein HuR, forms messenger ribonucleoprotein (mRNP) complexes with transiently expressed mRNAs encoding early-response transcription factors, including c-Fos, c-Jun, and Egr-1. Knockdown and overexpression assays demonstrated that proper posttranscriptional control of Egr-1 expression requires HuR-mediated translation control. Further analysis showed that the Egr-1 3'UTR, which contains AU-rich elements (AREs) and interacts directly with HuR, suppresses reporter gene expression and mediates posttranscriptional regulation of Egr-1 by HuR. These findings underscore an essential role for HuR in regulating early events during T cell activation.

Differential regulation of mRNA stability controls the transient expression of genes encoding Drosophila antimicrobial peptide with distinct immune response characteristics.

The tight regulation of transiently expressed antimicrobial peptides (AMPs) with a distinct antimicrobial spectrum and different expression kinetics contributes greatly to the properly regulated immune response for resistance to pathogens and for the maintenance of mutualistic microbiota in Drosophila. The important role of differential regulation of AMP expression at the posttranscriptional level needs to be elucidated. It was observed that the highly expressed Cecropin A1 (CecA1) mRNA encoding a broad antimicrobial spectrum AMP against both bacteria and fungi decayed more quickly than did the moderately expressed Diptericin mRNA encoding AMP against Gram negative bacteria. The mRNA stability of AMPs is differentially regulated and is attributed to the specific interaction between cis-acting ARE in 3'-UTR of AMP mRNA and the RNA destabilizing protein transactor Tis11 as shown in co-immunoprecipitation of the Tis11 RNP complex with CecA1 mRNA but not other AMP mRNA. The p38MAPK was further demonstrated to play a crucial role in stabilizing ARE-bearing mRNAs by inhibiting Tis11-mediated degradation in LPS induced AMP expression. This evidence suggests an evolutionarily conserved and functionally important molecular basis for and effective approach to exact control of AMP gene expression. These mechanisms thereby orchestrate a well balanced and dynamic antimicrobial spectrum of innate immunity to resist infection and maintain resident microbiota properly.

[The prevalence and risk factors for coronary stenosis in patients with cerebral infarction].

OBJECTIVE: To study the morbidity rate of and relevance to coronary stenosis in cerebral infarction patients. METHODS: CT coronary angiography was performed in 112 cases of cerebral infarction after CT cerebral angiography. Multivariate logistic regression analysis was carried out between the clinical data and coronary stenosis. RESULTS: In 112 cases receiving CT cerebral angiography, the morbidity rate of coronary stenosis was 46.4%. In 95 cerebral infarction patients, the morbidity rate of coronary stenosis was 51.6%. Multivariate logistic analysis showed that age, hypertension, hyperlipidemia, significant narrowing of cerebral artery were identified as independent predictors for coronary stenosis. CONCLUSIONS: Heart examination with 64 row CT should be routinely performed after cerebral angiography in cerebral infarction patients, especially in those with age greater than 65 years, hypertension, hyperlipidemia and significant narrowing of cerebral artery so as to detect coronary stenosis early.

[Observation on acupuncture at "Neitinggong" for antagonisting ototoxicity caused by gentamicin].

OBJECTIVE: To explore the effect of acupuncture at "Neitinggong" drug-induced deafness. METHODS: Guinea pig deafness model was prepared by injection of gentamicin (GM). Acupuncture was respectively given at the points "Neitinggong" "Tinggong" (SI 19) and non-acupoints on the auricle in the experimental animals in different groups and the effects of different points on the auditory brainstem response and cochlear hair cells were observed. RESULTS: There was a significant difference between GM group and Neitinggong group, and between GM group and Tinggong group. There was no significant difference between GM group and the auricle group, and between Neitinggong group and Tinggong group. CONCLUSION: Acupuncture at "Neitinggong" can strength the function of the internal ear, and relieve the injury of cochlear hair cells caused by gentamicin, which is an effective acupoint for treatment of drug-induced deafness.

[Large recombinant protein displayed on filamentous phage surface and its interaction with small molecule].

Recombinant proteins were expressed as fusions with the phagemid system of pHEN-KM13 and the characteristics and activities of the fusion proteins displayed on the surface of filamentous phagintain the were studiedon abilty. The altered titer of rescued phages from the phagemid system after trypsin treatment indicated the relative quantity of the phages displaying fusion proteins. The rescue phages displayed foreign proteins could keep the bacterial infection ability, while the bald phage without foreign protein displayed on its surface was sensitive to trypsin treatment and lost the bacterial infection ability. To determine the upper limit for filamentous phage display, four recombinant proteins, glutathione-S-transferase and glutathione-S-transferase fused with three various size peptide linkers, were fused to N terminus of capsid protein gp3 and rescued by helper phage KM13. The rescued phages which displayed fused protein with the size of 40kD or less maintain the infection ability. To assay the activity of the phage displayed protein, the known small molecule probe was used in the interaction study with protein incorporated on phage surface. Results showed that the glutathione-S-transferase on phage surface still bound to glutathione specifically. It indicated that the glutathione-S-transferase displayed on phage surface was correctly folded and functionally active. The results demonstrated that it was feasible to use small molecule probes to interact with the protein displayed on phage surface. In turn, the method described here also demonstrated that phage display system could be utilized to investigate the interactions between protein and small molecules.

Display of aggregation-prone ligand binding domain of human PPAR gamma on surface of bacteriophage lambda.

AIM: To display the aggregation-prone ligand binding domain (LBD) of the human peroxisome proliferator-activated receptor gamma (PPARgamma) on the surface of bacteriophages to establish an easy screening assay for the identification of PPARgamma ligands. METHODS: Plasmids were constructed for the expression of the PPARgamma LBD as a fusion to the N-terminus of the g3p protein of filamentous phage or the C-terminus of the capsid protein D (pD) of phage lambda. The fusion proteins were expressed in E coli and solubility characteristics were compared. Polyclonal antibodies against the LBD as well as the pD protein were prepared for Western blot analysis and phage capture assay. RESULTS: The pD-LBD fusion protein was partially soluble, whereas the LBD-g3p fusion protein was detected only in the insoluble fraction. The pD-LBD fusion protein was efficiently incorporated in phage particles. Furthermore, the LBD was shown to be displayed on the surface of bacteriophage lambda. On average, the pD-LBD fusion protein accounted for 28% of the total pD protein in the lambda head capsid. CONCLUSION: The hydrophobic PPARgamma LBD was expressed as a soluble form of fusion protein in E coli and displayed on the surface of bacteriophage lambda when it was fused to the lambda pD protein. The lambda pD fusion system could be used for improving the solubility of proteins that tend to form inclusion bodies when expressed in E coli. The lambda phage particles displaying the LBD of PPARgamma may be of great value for the identification of novel PPARgamma ligands.

Elucidation of subfamily segregation and intramolecular coevolution of the olfactomedin-like proteins by comprehensive phylogenetic analysis and gene expression pattern assessment.

The categorization of genes by structural distinctions relevant to biological characteristics is very important for understanding of gene functions and predicting functional implications of uncharacterized genes. It was absolutely necessary to deploy an effective and efficient strategy to deal with the complexity of the large olfactomedin-like (OLF) gene family sharing sequence similarity but playing diversified roles in many important biological processes, as the simple highest-hit homology analysis gave incomprehensive results and led to inappropriate annotation for some uncharacterized OLF members. In light of evolutionary information that may facilitate the classification of the OLF family and proper association of novel OLF genes with characterized homologs, we performed phylogenetic analysis on all 116 OLF proteins currently available, including two novel members cloned by our group. The OLF family segregated into seven subfamilies and members with similar domain compositions or functional properties all fell into relevant subfamilies. Furthermore, our Northern blot analysis and previous studies revealed that the typical human OLF members in each subfamily exhibited tissue-specific expression patterns, which in turn supported the segregation of the OLF subfamilies with functional divergence. Interestingly, the phylogenetic tree topology for the OLF domains alone was almost identical with that of the full-length tree representing the unique phylogenetic feature of full-length OLF proteins and their particular domain compositions. Moreover, each of the major functional domains of OLF proteins kept the same phylogenetic feature in defining similar topology of the tree. It indicates that the OLF domain and the various domains in flanking non-OLF regions have coevolved and are likely to be functionally interdependent. Expanded by a plausible gene duplication and domain couplings scenario, the OLF family comprises seven evolutionarily and functionally distinct subfamilies, in which each member shares similar structural and functional characteristics including the composition of coevolved and interdependent domains. The phylogenetically classified and preliminarily assessed subfamily framework may greatly facilitate the studying on the OLF proteins. Furthermore, it also demonstrated a feasible and reliable strategy to categorize novel genes and predict the functional implications of uncharacterized proteins based on the comprehensive phylogenetic classification of the subfamilies and their relevance to preliminary functional characteristics.

The existence of a putative regulatory element in 3'-untranslated region of proto-oncogene HOX11's mRNA.

HOX11 encodes a homeodomain-containing transcription factor which directs the development of the spleen during embryogenesis. While HOX11 expression is normally silenced through an unknown mechanism in all tissues by adulthood, the deregulation of HOX11 expression is associated with leukemia, such as T-cell acute lymphoblastic leukemia. The elucidation of regulatory elements contributing to the molecular mechanism underlying the regulation of HOX11 gene expression is of great importance. Previous reports of HOX11 regulatory elements mainly focused on the 5\'-flanking region of HOX11 on the chromosome related to transcriptional control. To expand the search of putative cis-elements involved in HOX11 regulation at the post-transcriptional level, we analyzed HOX11 mRNA 3\'-untranslated region (3\'UTR) and found an AU-rich region. To characterize this AU-rich region, in vitro analysis of HOX11 mRNA 3\'UTR was performed with human RNAbinding protein HuR, which interacts with AU-rich element (ARE) existing in the 3\'UTR of many growth factors' and cytokines' mRNAs. Our results showed that the HOX11 mRNA 3\'UTR can specifically bind with human HuR protein in vitro. This specific binding could be competed effectively by typical ARE containing RNA. After the deletion of the AU-rich region present in the HOX11 mRNA 3\'UTR, the interaction of HOX11 mRNA 3\'UTR with HuR protein was abolished. These findings suggest that HOX11 mRNA 3\'UTR contains cis-acting element which shares similarity in the action pattern with ARE-HuR interactions and may involve in the posttranscriptional regulation of the HOX11 gene.

hOLF44, a secreted glycoprotein with distinct expression pattern, belongs to an uncharacterized olfactomedin-like subfamily newly identified by phylogenetic analysis.

Secreted proteins are indispensable for the development and differentiation of multicellular organisms. Cloning and characterization of novel or hypothetical genes encoding these proteins are therefore inviting great incentives. Using bioinformatics tools and experimental approaches, we isolated and characterized a human secreted glycoprotein, hOLF44, which contains a highly conserved olfactomedin-like (OLF) domain in the C-terminal. However, phylogenetic analysis revealed that hOLF44 is not clustered into any of the OLF subfamilies containing characterized members, and obviously falls into a newly identified uncharacterized OLF subfamily. Western blot analysis showed that hOLF44 protein is robustly secreted from the transfected COS-7 cells. Expression levels of hOLF44 mRNA are abundant in placenta, moderate in liver and heart, whereas fairly weak in other tissues examined. Immunohistochemical study on human term placenta demonstrated that hOLF44 is mainly localized extracellularly surrounding the syncytiotrophoblastic cells and very rarely expressed in the maternal decidua layer. These results suggest that hOLF44 may have matrix-related function involved in human placental and embryonic development, or play a similar role in other physiological processes. The further functional characterization of hOLF44 may provide insights into a better understanding of the newly identified OLF subfamily.

A modified quantitative EMSA and its application in the study of RNA--protein interactions.

Electrophoretic mobility shift assay (EMSA) is used to detect the complex of protein and nonradioisotope-labeled probe qualitatively. In this paper, we describe a modified quantitative EMSA, which uses biotin-labeled RNA in the complex formation. The RNA-protein complex is separated by agarose gel electrophoresis and capillary transferred to a positively charged nylon membrane. It is then detected through a secondary detection system using 5-bromo-4-chloro-3-indolyl phosphate (BCIP)-nitroblue tetrazolium (NBT) as the colorimetric precipitating substrate. After scanning and quantification by an image analysis program, ImageQuant, it was observed that the optical density of the bands was proportional to the decadic logarithm value of standard RNA quantities in the tested range. By using the standard curve of the optical densities plotted against the logarithm values of RNA quantities in the linear range, we can calculate RNA quantities according to the optical density of the band and make a quantitative analysis of EMSA. This method was applied successfully in the study of the interactions between the AU-rich element (ARE) and HuR, which is one of the human members of the (embryonic lethal abnormal vision) ELAV family. The results reveal that the biotin-labeled AUFL transcripts in the RNA-protein complex can be detected quantitatively, while maintaining the biological features of its binding ability to the HuR protein. By this method, it is possible to conduct qualitative and quantitative analyses of the EMSA in the study of RNA-protein interactions.

The existence of a putative post-transcriptional regulatory element in 3'-UTR of Drosophila antibacterial peptide diptericin's mRNA.

Antibacterial peptides' genes are rapidly and transiently expressed on immune stimulation, which is the characteristic of immediate early genes. It implies post-transcriptional regulation is an important pathway in antibacterial peptides' gene expression. In a search of putative post-transcriptional regulatory elements, we found a segment of an AU-rich sequence in 3'-untranslated region (UTR) of drosophila diptericin mRNA. 3'-UTR of diptericin mRNA can be specifically bound with Elav and this binding can be competed with the typical AU-rich element (ARE) of c-fos mRNA. These results suggest that the AU-rich sequence in the 3'-UTR of diptericin mRNA may be a cis-acting element and involved in post-transcriptional regulation.

Cloning and characterization of a novel human secretory protein: secretogranin III.

We have cloned a new member of the granin family, termed human secretogranin III (SgIII), that encodes 468 amino acid residues. The human SgIII protein possesses an N-terminal signal peptide, seven dibasic sites, and the repeated DSTK sequences. These structure characteristics are similar to other members of secretogranin family. The human SgIII has homologous proteins in mouse, rat, and Xenopus laevis. Genomic organization shows the gene includes 12 coding exons spanning 39 kb of genomic DNA on the human chromosome 15. Human SgIII is expressed widely as showed in Northern blot and its cDNA hybridizes to 2.2 kb and 1.9 kb bands in many tissues, with two additional 4.5 kb and 3.3 kb bands in brain. Subcelluar localization and immunoblotting indicated SgIII was secreted out of cell through trans-Golgi network (TGN). SgIII may take effect in the biogenesis of secretory granules as a helper protein and be involved in the production or release of peptide hormones in the regulated secretory pathway.