Molecular cloning, expression and functional characterization of tumor necrosis factor (TNF) receptor-associated factor (TRAF)-interacting protein (TRIP) in grass carp, Ctenopharyngodon idella.

TRIP (Tumor Necrosis Factor (TNF) Receptor-Associated Factor (TRAF)-Interacting Protein), a member of the TNF superfamily, plays a crucial role in the modulation of inflammation in vertebrates. However, no information about TRIP is available in teleosts. In this study, the full-length cDNA of TRIP, containing a 5'UTR of 112 bp, an ORF of 1359 bp, and a 3'UTR of 29 bp before the poly (A) tail, was cloned from grass carp, Ctenopharyngodon idella. The TRIP gene encoded a protein of 452 amino acids with an estimated molecular mass of 51.06 KD and a predicted theoretical isoelectric point (pI) of 9.11. Quantitative real-time PCR analysis revealed that TRIP mRNA was expressed in all the tissues examined in grass carp, with the highest expression in the kidney, followed by the intestine and thymus. However, lower levels of expression were also detected in fat, spleen, liver, gonad and heart. Subcellular localization and two-hybrid analysis revealed that TRIP was located in the nucleus and that it interacted with TRAF1 and TRAF2 in HEK293T cells. Furthermore, similar to TNF-α, IL-10 and TRIP mRNA expression was upregulated in the spleen of fish fed high-fat or high-carbohydrate diets, suggesting that TRIP might be associated with the response to excessive energy intake. The mRNA relative expression of TRIP was significantly reduced (P < 0.05) after hepatocyte of C. idella was treated with 2 µg/mL lipopolysaccharide (LPS) for 4 h, while the expression levels of inflammatory cytokines TNF-α and IL-10 were significantly increased (P < 0.05). Taken together, these results indicate that TRIP might play important roles in immune defense and has the potential to be used as a anti-inflammation target in grass carp.

PGC-1ß modulates the expression of genes involved in mitochondrial function and adipogenesis during preadipocyte differentiation.

This study determined the expression of genes involved in mitochondrial function and adipogenesis at mRNA and protein levels by transfecting rat differentiating preadipocytes with siRNA/Lipofectamine complex and pcDNA-PGC-1ß (peroxisome proliferator-activated receptor-γ coactivator-1ß)/Lipofectamine complex, respectively, to further elucidate the role of PGC-1ß in white preadipocyte differentiation. The results showed that the transfection of PGC-1ß siRNA inhibited the expressions of mitochondrial genes malate dehydrogenase, carnitine palmitoyltransferase 1, nuclear respiratory factor 1, ATP synthesis, adipocyte differentiation key transcription factor peroxisome proliferator-activated receptor-γ, sterol regulatory element binding protein 1c and fatty acid synthetase, whereas the triglyceride synthesis was retarded (p < 0.05). Furthermore, overexpression of PGC-1ß up-regulated the expressions of adipogenic and mitochondrial biosynthetic marker genes and promoted triglyceride accumulation during 3T3-L1 adipocyte differentiation. These observations suggest that PGC-1ß modulates the expression of mitochondrial function and adipogenesis-related genes and affects white preadipocyte differentiation.

[Meta-analysis of the effects of supplemental parenteral nutrition on prognosis of critically ill patients].

Objective: To compare the effects of supplemental parenteral nutrition (SPN) and enteral nutrition (EN) on prognosis of critically ill patients in intensive care unit (ICU) using meta-analysis. Methods: Foreign language databases including PubMed, Embase, Cochrane Library, and Cochrane Central Register of Controlled Trials were retrieved with the search terms of " supplemental parenteral nutrition, parenteral nutrition, enteral nutrition, critically ill" , and Chinese database SinoMed database was retrieved with the search terms of ",,," to obtain the publicly published randomized controlled trials about the effects of SPN and EN supportive treatment on prognosis of critically ill patients in ICU from the establishment of each database to December 2018. The Google Scholar was retrieved for supplement. The outcome indexes included the infection rate, anti-infection time, antibiotic-free time, ICU overall mortality, overall mortality during hospitalization, mechanical ventilation time, length of ICU stay, and length of hospital stay. RevMan 5.3 and Stata 14.0 statistical software were used to conduct a meta-analysis of eligible studies. Results: A total of 794 patients were included in 8 studies, including 387 patients in SPN group who received SPN and EN and 407 patients in EN group who only received EN. The bias risks of the eight studies included were uncertain. Compared with that of EN group, the infection rate of patients in SPN group was significantly decreased (relative risk=0.79, 95% confidence interval=0.66-0.94, P<0.01). However, there were no statistically significant differences in ICU overall mortality, overall mortality during hospitalization, mechanical ventilation time, length of ICU stay, and length of hospital stay of patients between SPN group and EN group. The subgroup analysis showed that the risks of bias of studies and follow-up time might be sources of the heterogeneity of mechanical ventilation time. There was publication bias in ICU overall mortality (P<0.05), while no publication bias in the other outcome indexes (P>0.05). Conclusions: SPN supportive treatment can decrease the infection rate of critically ill patients in ICU, but it has no obvious influences on overall mortality, mechanical ventilation time, and length of hospital stay.

Hepatocellular carcinoma cells containing hepatitis B virus X protein have enhanced invasive potential conditionally.

AIMS: To establish a sustaining hepatitis B virus X protein expressed Chang liver cell line and to explore their biological behaviours of invasive potential induced by hepatitis B virus X protein. METHODS: Polymerase chain reaction was used to amplify the HBx gene from the whole hepatitis B virus genome. The gene was then subcloned into the eukaryotic expression vector pcDNA3.1 to construct the pcDNA3.1-HBx plasmid. Gene transfection mediated by Lipofectamine was used to introduce the plasmid into the human liver cell line Chang, and stable expression of the HBx gene was detected. RESULTS: HBx gene was cloned from the transfected Chang liver cells by reverse transcription-polymerase chain reaction, and confirmed by electrophoresis. The stably transfected Chang cells expressing HBx with malignant characteristics were verified and compared with control cells in terms of their growth curves, clonogenicity, wound healing abilities, migration and metastasis. CONCLUSION: The stabilising human liver cell lines Chang liver containing HBx gene expression have been established successfully. The invasive potential of Chang cells was conditionally enhanced by HBx transfection.

Evaluation of echinacea for treatment of the common cold.

Considered to have immunostimulating activity, echinacea is a widely used phytomedicinal for treatment of the common cold and upper respiratory tract infections (URTIs). We reviewed the literature from the MEDLINE database (January 1966-July 1999), International Pharmaceutical Abstracts (IPA) online database, Cambridge Scientific Abstracts Biological Sciences online database, Alt-Health Watch online database, EMBase CD-ROM database, and references from published articles, reviews, and letters to evaluate evidence from clinical trials of echinacea's purported efficacy for treating or preventing URTIs. Twelve clinical studies published from 1961-1997 concluded that echinacea was efficacious for treating the common cold, but the results are unclear due to inherent flaws in study design. Five trials were published since 1997; two showed that echinacea lacked efficacy for treating and preventing URTI symptoms, and three concluded that it was effective in reducing the frequency, duration, and severity of common cold symptoms. Again, these results are unclear because of methodologic uncertainties, such as small populations and use of noncommercially available, nonstandardized dosage forms. Although evidence for echinacea's efficacy is inconclusive, it appears to be safe. Patients without contraindications to it may not be dissuaded from using an appropriate preparation to treat the common cold.

Pharmacists' knowledge and attitudes toward herbal medicine.

OBJECTIVE: The use and sales of herbal medications have increased dramatically over the past several years. Pharmacists are in an ideal position to educate patients about herbal medicines. This study was intended to determine the knowledge and attitudes of pharmacists regarding herbal medications. METHODS: A survey was distributed to pharmacists at several state and regional meetings in Virginia and North Carolina between August and October 1998. The survey evaluated demographic data, attitudinal scales, and a 15-item herbal medicine knowledge test. Pharmacists immediately returned the surveys to the distributor on completion. RESULTS: Of the 217 surveys distributed, 164 met the inclusion criteria for further evaluation. Of the pharmacists surveyed, 68.0% practiced in a community pharmacy, 45.1% had previous continuing education on herbal medications, and 73.6% sold herbal medications in their practice settings. The average score on the herbal knowledge test was 6.3 (maximum score of 15). Pharmacists with previous continuing education scored significantly higher than those without prior continuing education (p < 0.001). Of the 15 questions, the five that pharmacists were most likely to answer correctly assessed the uses of herbal medications. Additionally, pharmacists with prior continuing education or with access to herbal medication information at their practice site were more likely to agree that providing information about herbal medication is a pharmacist's professional responsibility (p = 0.02 and p = 0.01, respectively). CONCLUSIONS: The findings from this study demonstrate that pharmacists were more likely to answer correctly about the uses of herbal medications than about drug interactions, adverse drug effects, and precautions of herbal medications. Additionally, pharmacists with previous continuing education on herbal medications were more knowledgeable about these products. With the increasing use of herbal medications, there is a greater need for pharmacy training programs in this area.