Temporary Ischemia Time Before Snap Freezing Is Important for Maintaining High-Integrity RNA in Hepatocellular Carcinoma Tissues.

Background: High-throughput transcript sequencing plays an important role in the study of hepatocellular carcinoma (HCC) occurrence and development. High-quality biospecimens, especially high-quality RNA, are the most basic prerequisites for obtaining good transcript sequencing data. Our purpose was to explore the treatment conditions of in vitro ischemic tissue samples that can be used to obtain high-integrity RNA before freezing the samples in liquid nitrogen. Materials and Methods: Postoperative tumor tissues (T) and adjacent normal tissues (AN) from 50 HCC patients were randomly selected from May 5, 2017, to June 15, 2017. Postoperative tissue specimens from each HCC patient were stratified by tissue type (T or AN), ischemia time (minutes), and ischemia temperature (°C) into 16 groups: T-4°C-15 minutes, T-4°C-30 minutes, T-4°C-60 minutes, T-4°C-120 minutes, T-24°C-15 minutes, T-24°C-30 minutes, T-24°C-60 minutes, T-24°C-120 minutes, AN-4°C-15 minutes, AN-4°C-30 minutes, AN-4°C-60 minutes, AN-4°C-120 minutes, AN-24°C-15 minutes, AN-24°C-30 minutes, AN-24°C-60 minutes, and AN-24°C-120 minutes. RNA integrity was detected by RNA integrity number (RIN) and 1% agarose gel electrophoresis. Results: At an ischemia temperature of 4°C and ischemia time of >30 minutes, the RIN of T began to decrease. RIN also gradually decreased in T at an ischemia temperature of 4°C and in both T and AN at an ischemia temperature of 24°C for ischemia times 15, 30, 60, and 120 minutes. For an ischemia time ≤15 minutes and ischemia temperature 4°C or 24°C, the RINs of T and AN were significantly different. Furthermore, at ischemia temperature 4°C and ischemia time 30 and 60 minutes or ischemia temperature 24°C and ischemia time 30 minutes, the RIN of T was higher compared with AN. However, there was no significant difference in RIN between T and AN under other treatment conditions. Conclusions: Tissue quality is adversely affected by ischemia time and ischemia temperature. Therefore, temporary ischemia time (≤15 minutes) before snap freezing is key for maintaining high-integrity RNA in HCC tissues.

[Senile habitual constipation treated with auricular therapy based on the pattern/syndrome differentiation: a randomized controlled trial].

OBJECTIVE: To compare the efficacy on senile habitual constipation between the auricular therapy based on the pattern/syndrome differentiation and the conventional auricular point sticking therapy. METHODS: Two-hundred cases were randomized into a group of the auricular therapy based on the pattern/syndrome differentiation (an auricular differentiation group) and a group of conventional auricular point sticking therapy (an auricular sticking group), 100 cases in each one. In the auricular differentiation group, Brain Stem (AT3,4i), Occiput (AT3), Subcortex (AT4), Large Intestine (CO7), Triple Energizer (CO17), Abdomen (AH8), Endocrine (CO18) and Constipation Point were taken as the main points. According to the pattern/syndrome differentiation, for excessive heat in the stomach and intestine, Stomach (CO4) and Small Intestine (CO6) were added; for blockage of lung qi, Lung (CO14) was added; for spleen and kidney deficiency, Spleen (CO13) and Kidney (CO10) were added. In the auricular sticking group, CO7, Rectum (HX2), Sympathetic Nerve (AH6a) and AT4 were selected. In both groups, the patients were required to press the points four times a day by themselves. The auricular points were changed once every 3 to 4 days, and the two ears were alternated. The eight treatments made one session and two sessions were required totally. The clinical efficacy and the changes of the score for the quality of life before and after treatment were observed in the two groups. RESULTS: The effective rate was 92.0% (92/100) in the auricular differentiation group, which was superior to 76.0% (76/100) in the auricular sticking group (P < 0.05). The score for the quality of life after treatment was reduced to different extents for the patients in the two groups (both P < 0.05). The score decrease in the auricular differentiation group was much more apparent as compared with that in the auricular sticking group (P < 0.05). CONCLUSION: The auricular therapy based on the pattern/syndrome differentiation is safe and effective in the treatment of senile habitual constipation and its efficacy is superior to the conventional auricular point sticking therapy.