Cold-inducible ribonucleic acid-binding protein attenuates acute kidney injuries after deep hypothermic circulatory arrest in rats.
The cold-inducible ribonucular acid binding protein (CIRP) has been identified to play a role in the antipoptotic effect of hypothermia. We sought to investigate CIRP renoprotection in a rat model of a deep hypothermic circulatory arrest. The overexpression and reversing of the CIRP have been obtained in vivo by directly injuring the vectors of lentivirus containing lentivirus lentivirus (PL) / of the internal fluorescent protein / green fluorescent protein (GFP) -CIRP or hairdressing RNA PL / short) / FF-FREE Inductible RNA binding protein (F-CIRP) -a in the renal parenchyma of 7-day rats before the deep hypothermic circulatory arrest under ultrasound guidance.
The vehicles or vectors of control lentiviruses were given to the control group or the control vector group, respectively. The renal function and activity of apoptosis have been evaluated by serum Cystatin C, the lipocalin of serum gelatinasse / gelatinasse and the deoxynucleotidine terminal of gelatinneity of the gelatinque 2′-deoxyudidine, the dosage of the Nick-end labeling of 5′-triphosphate at 24 hours after surgery. The expression of CIRP messenger RNA (mRNA) was evaluated by a quantitative real-time polymerase chain reaction. The expression of CIRP proteins and Caspase 3 has been tested by Western blot.
Compared to the SHAM Group, the Rats of the control group showed an increased expression of CIRP mRNA, CIRP protein, Caspase 3 and apoptotic frequency (p <0.01). However, in relation to the control group, the Rats of the PL / IRES / GFP-CIRP Group showed a significant decrease in CASPASE 3 activities and apoptosis, while increasing the increased expression of mRNA and protein. CIRP. Rats in the PL / SHRNA / F-CIRP-A group showed increased activities of CASPASE 3 and apoptosis and a decrease in a further decrease in CRNA and CIRP protein (p <0, 01), compared to the control group. The renal function was particularly protected in the PL / IRES / GFP-CIRP group and altered in the PL / SHRNA / F-CIRP-A group.
Our results suggest that the CIRP exerts a robust renoprotective effect by inhibiting apoptosis in the rat model of a deep hypothermic circulatory arrest.
Storage of long-term room temperature of dry ribonucleic acid to be used in the ARN-SEQ analysis.
RNA is an essential biological material for genomics and translational medicine research. As such, its storage of Biobanking is an important field of study. Traditionally, long-term storage in the cold (usually freezers or liquid nitrogen) is used to maintain high quality RNA (in terms of quantity and integrity). The conservation of the ambient temperature (RT) provides a cold alternative, which is in the grip of serious problems (mainly costs and security), for the long-term storage of RNA. In this study, we evaluated the performance of several RT storage procedures, including the Imagene’s RNASHELL®, where RNA is dried and maintained protected from the atmosphere and vacuum drying of RNA with such additives. that the Imagene stabilization solution and a house -made of trehalose solution.
This evaluation was completed by accelerated (equivalent to 10 years for Rnasell) and real-time studies (4 years). To verify the quality and integrity of RNA, we used RNA and RNA Integrity Number Values. Our study shows that the isolation of the atmosphere offers a higher protective effect for the storage of RNA with respect to vacuum drying and demonstrates that Rnasell allows a satisfactory RNA quality for long-term RT storage. Thus, the quality of the RNA could meet the demand for downstream applications such as RNA-SEQ.
Cold-inducible ribonucleic acid-binding protein attenuates acute kidney injuries after deep hypothermic circulatory arrest in rats.
HASHINGDOWN OF IL-6 ribonucleic acid interference improves the effectiveness of cisplatin in laryngeal cancer stem cells by regulating the regulation path of the IL-6 / STAT3 / HIF1 regulatory channel.
CONTEXT
Cisplatin has been used in the treatment of many cancers, including larynx cancer; However, its effectiveness can be reduced due to the development of drug resistance. This study was intended to determine whether the removal of interleukin-6 (IL-6) can enhance the efficacy of cisplatin in larynx cancer stem cells (CSC) and the potential involvement of the signal transducer and the activator of transcription 3 (STAT3) and the hypoxia 1 inducible factor (HIF1) for this purpose.
Methods
The sorting cells of the fluorescence activated cells were identified in the sorting cells activated by fluorescence. The mRNA and IL-6, STAT3 and HIF1 phrases were examined with respectively the reaction of the polymerase chain in real time and the western ball. Cell proliferation was measured by MTT dosage. Turmoricity was measured by a colony formation dosage and an invasion was determined by a cell invasion test. Apoptotic cells have been counted by flow cytometry. Immunohistochemistry was performed to detect immunoreactive IL-6, STAT3 and HIF1 cells in xenograft.
RESULTS
IL-6, STAT3 and HIF1 mRNA and protein levels have been significantly increased in HEP2-CSC with respect to those of HEP2 cells. The application of SIRNA-IL-6 to Hashdown IL-6 resulted in a significant decrease in IL-6, STAT3 and HIF1 mRNA and protein rates. IL-6 HASHINGDOWN reduced cell proliferation, tumorigenetness and invasion and increased apoptosis within CSC. Improved deletion degrees of these parameters were observed when the IL-6 return was associated with cisplatin in these CSCs. The results of the Xenograft study showed that the combination of reversal and cisplatin IL-6 inhibited the growth of xenografts with respect to that obtained in the single injected group of cisplatin.
Description: EcoPURE Total RNA Kit is designed as a simple and convenient purification of high quality total RNA including small RNAs (e.g. microRNAs) from whole blood, cultured cells, and frozen or fresh tissues. This kit utilizes chaotropic ions and silica-based membrane technology, eliminating the need for expensive resins, hazardous phenol-chloroform extractions, β-mercaptoethanol, or time-consuming alcohol precipitation. The standard protocol lasts less than 10 minutes at room temperature and purified RNA can be effectively used in routine downstream applications including cDNA synthesis, northern blotting, differential display, primer extension, and mRNA selection.
Immunoreactive IL-6, STAT3 and HIF1 Cell numbers were significantly reduced in the tumor tissues of IL-6. IL-6, STAT3 and HIF1 Immunoreactive cell accounts have been further reduced in the tissues where the overturning IL-6 has been combined with cisplatin treatment with respect to fabric cisplatin only.
