PCDH10 inhibits invasion of lymphoma cells by regulating ß-catenin.

OBJECTIVE: The aim of this study was to investigate the effect of protocadherin 10 (PCDH10) on the invasive potential of lymphoma cells by regulating matrix metalloproteinase-7 (MMP7) and MMP9 via targeting ß-catenin. MATERIALS AND METHODS: The mRNA and protein expressions of PCDH10, ß-catenin, MMP7 and MMP9 in lymphoma cell lines were examined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Raji cells with low expression of PCDH10 were transiently transfected with pCMV5-HA-PCDH10 or pcDNA5-His-ß-catenin. Meanwhile, HUT-78 cells with high expression of PCDH10 were transfected with PCDH10-shRNA or ß-catenin-shRNA. Subsequently, the expression levels of ß-catenin, MMP7 and MMP9 in transfected lymphoma cells were determined as well. In addition, the regulatory effects of PCDH10 on the invasive potential of lymphoma cells were explored by transwell assay. RESULTS: PCDH10 expression was negatively correlated with the expressions of ß-catenin, MMP7 and MMP9 in several lymphoma cell lines. Transfection of HA-PCDH10 in human-derived malignant B lymphoma cell line Raji markedly down-regulated the protein levels of ß-catenin, MMP7 and MMP9. However, the mRNA level of ß-catenin was not influenced by PCDH10. Interference with PCDH10 in HUT-78 cells with high expression of PCDH10 significantly increased the protein expressions of ß-catenin, MMP7, and MMP9. However, no significant changes were observed in the mRNA expression of ß-catenin. In addition, knockdown of ß-catenin in cells with high expression of PCDH10 remarkably down-regulated the expression levels of MMP7 and MMP9. CONCLUSIONS: PCDH10 overexpression in lymphoma cells downregulates ß-catenin expression, as well as inhibits the expressions of MMP7 and MMP9, eventually inhibiting the invasive potential of lymphoma cells.

Value of Model for End-Stage Liver Disease-Serum Sodium Scores in Predicting Complication Severity Grades After Liver Transplantation for Acute-on-chronic Liver Failure.

BACKGROUND: Preoperative evaluation is extremely important for patients undergoing liver transplantation (LT) for acute-on-chronic liver failure (ACLF). It is unclear that whether preoperative Model for End-Stage Liver Disease-Serum Sodium (MELD-Na) score has a decisive effect on the complication grade after LF for ACLF. This study is aimed to explore the value of preoperative MELD-Na scores in predicting complication severity grades post LT for ACLF. METHODS: One hundred fifty-nine patients enrolled in the study who underwent LT for ACLF between August 1, 2004, and September 1, 2014, were retrospectively analyzed. The Accordion Severity Grading system was adopted to classify the complication severity grade post LT: Grade 1 (mild), grade 2 (moderate), grades 3-5 (severe), and grade 6 (death). The area under the curve was calculated by plotting the receiver operating characteristic curve for evaluating the diagnostic accuracy of MELD-Na score for severe grade and mortality after LT. The correlation between MELD-Na score with complication severity grade post LT was demonstrated by Spearman correlation and multivariate analysis. The MELD-Na based nomogram was constructed to predict short-term mortality (grade 6). RESULTS: The incidences of postoperative complications at all grade levels were: grade 2: 43 patients (27.0%, MELD-Na 27.3 ± 7.4), grade 3: 41 patients (25.8%, MELD-Na 32.7 ± 12.4), grade 4: 31 patients (19.5%, MELD-Na 34.3 ± 12.1), grade 5: 9 patients (5.7%, MELD-Na 30.7 ± 12.3), grade 6: 35 patients (22%, MELD-Na 37.1 ± 10.4). There was no grade 1 patient. The area under the curve of the MELD-Na scores for severe and death group were 0.631 (P < .05, 95% confidence interval [CI], 0.533-0.728) and 0.670 (P < .05, 95% CI, 0.574-0.766) respectively. The MELD-Na score was significantly correlated with the Accordion Severity Grade (rho 0.297, P < .01) by Spearman correlation analysis. Multivariate analysis confirmed that a MELD-Na score ≥ 25 was the only risk factor for postoperative severe grade complications (P < .05, odds ratio = 4.35) and that MELD-Na ≥ 35 was one risk factor for postoperative mortality (P < .01, hazard ratio = 4.72). MELD-Na ≥ 35 combined with other parameters (female, age, systematic infection, and intraoperative placement of the T-tube) in a constructed nomogram model had a good calibration curve with C-concordance of 0.790. CONCLUSIONS: MELD-Na scores are significantly correlated with Accordion Severity Grades. It can effectively predict the complication severity grade after LT for ACLF.

Conjugated linoleic acid synthesis-related protein proteasome subunit α 5 (PSMA5) is increased by vaccenic acid treatment in goat mammary tissue.

This study was conducted to identify proteins associated with the endogenous synthesis of conjugated linoleic acid (CLA) from trans-vaccenic acid (TVA; trans-11 C18:1, a precursor for CLA endogenous synthesis) in mammary tissues. Six lactating goats were divided into 2 groups. One group was given an intravenous bolus injection of TVA (150mg) twice daily over 4 d; the other group received saline injections. Treatment with TVA increased the concentration of cis-9,trans-11 CLA and TVA in goat milk. Additionally, TVA treatment increased the expression of stearoyl-CoA desaturase (SCD) in mammary tissue. Using 2-dimensional gel electrophoresis and electrospray ionization quadrupole time-of-flight mass spectrometry, 3 proteins affected by infusions of TVA were identified. Proteasome (prosome, macropain) subunit α type 5 (PSMA5) was upregulated, whereas peroxiredoxin-1 and translationally controlled tumor protein 1 were downregulated in TVA-treated animals compared with the vehicle-injected controls. Only the effect of TVA on PSMA5 could be confirmed by Western blot analysis. To further explore the regulation of PSMA5 in mammary epithelial cells when TVA is converted into CLA, we used a differentiated bovine mammary epithelial cell line treated with TVA for 6h. Changes in cis-9,trans-11 CLA concentrations and mRNA expression patterns of both SCD and PSMA5 were monitored. The concentration of cis-9,trans-11 CLA increased after TVA treatment. The mRNA expression level of PSMA5 was significantly elevated to 6h, but SCD mRNA expression only increased in 2h after TVA treatment. These results indicate that PSMA5 is highly expressed in goat mammary tissue and bovine mammary epithelial cells when TVA is converted into CLA. Our data suggest that PSMA5 protein is associated with CLA biosynthesis in mammary tissue.

Transforming activity of retroviral genomes encoding Gag-Axl fusion proteins.

Retroviral genomes encoding a portion of the Moloney murine leukemia virus Gag protein fused to portions of the murine axl cDNA were constructed so as to mimic naturally occurring transforming viruses. Virus MA1 retained 5 amino acids of the extracellular domain and the complete transmembrane and intracellular domains of Axl; virus MA2 retained only the intracellular Axl sequences beginning 33 amino acids downstream of the transmembrane region. Although both viruses could transform NIH 3T3 cells, they induced different morphological changes. MA1 transformants became elongated and assumed a cross-hatched pattern, while MA2 transformants were round and very refractile and grew to high density. Gag-Axl and Glyco-Gag-Axl proteins were detected in both types of transformed cells and were predominantly localized to the cytoplasmic compartment. When cell-free v-axl virus supernatants were introduced into wild-type BALB/c neonates, Rag-2-deficient mice, or c-myc transgenic mice, they did not cause tumors in a 3-month period. However, MA2-transformed NIH 3T3 cells, but not MA1 or control cells, could establish sarcomas by subcutaneous or intraperitoneal injection into BALB/c neonates. These results show that the transforming potential of the axl gene can be activated by truncation of the extracellular domain of the receptor and fusion of the remaining sequence to the gag gene.

Effects of a new cholinolytic drug of tropanes and its optical isomers on central muscarinic and nicotinic receptors.

2 alpha-(2',2'-disubstituted-2'-hydroxy-ethoxy)tropane (2 alpha-DHET cholinolytic, is a racemic tertiary amine with two chiral carbonic atoms. It has four optical isomers whose absolute configurations are 1R-2 alpha-2'S, 1R-2 alpha-2'R 1S-2 alpha'R and 1S-2 alpha-2'S. These compounds showed both antimuscarinic and antinicotinic activity, blocking both muscarinic and nicotinic receptors. Central muscarinic receptors, rather than nicotinic receptors, have a stereoselective specificity to these compounds. The 2'R configurations are more suitable to the stereostructure of the binding site of muscarinic receptors than the 2'S configuration.

[Studies on the stereochemistry and structure-activity relationship of cholinolytic compounds 3-(2-phenyl-2-cyclopentyl-2-hydroxyl-ethoxy)-quinuclidines].

Four optical isomers of the new cholinolytic compound 3-(2-phenyl-2-cyclopentyl-2-hydroxyl-ethoxy)-quinuclidine (I) have been asymmetrically synthesized by two methods. Method one: Recemic 1-phenyl-1-cyclopentylepoxyethane reacting with 3R or 3S-quinuclidinol produces a mixture of (R-1) and (R-2) or (S-1) and (S-2) respectively. The chemical yields varied from 57% to 78%. The highest % de is 22 and the major product is (R-1) or (S-1). Method two: Grignard reaction of 3R or 3S-benzoyl-methoxy-quinuclidine with cyclopentyl magnesium bromide yields a mixture of (R-1) and (R-2) or (S-1) and (S-2). The chemical yield is 80%. The highest % de is 81 and the major product is (R-2) or (S-2). Preliminary evaluation of the four new optical isomers revealed the following series of biological potencies: (R-2) greater than (I) greater than (S-1) greater than (R-1) greater than (S-2). In the coupling of the compounds with the active centers of M receptors, the absolute configurations in carbon-3 of the quinuclidinyl group and carbon-2 of the substituted ethyl group play an important role. The influence of carbon-2R is greater than that of carbon-3R on cholinolytic potency.

[1H, 13C NMR and stereochemistry studies of 3S-substituted alkoxylquinuclidine by two-dimensional and NOE difference NMR techniques].

By using optically pure 3S-quinuclidinol, two diastereoisomers (3S-1) and (3S-2) of 3-(substituted) alkoxy-quinuclidine were synthesized. 1H and 13C NMR spectra of (3S-1) and (3S-2) have been completely analyzed utilizing double-quantum filtered COSY, 13C-1H COSY and NOE difference experiment. The NOE difference experiment was used to determine the absolute configuration at C-11 of the diastereomers. According to the results of NOE difference and variable concentration NMR experiments, the configuration about C-11 of (3S-1) is designated as R (1A), whereas the configuration about C-11 of (3S-2) is designated as S (1B). The result was confirmed by X-ray diffraction analysis.