Contribution of Borneolum syntheticum to the Intervention Effect of Liuwei Dihuang Pill () on Experimental Retinal Degeneration.

OBJECTIVE: To observe the contribution of Borneolum syntheticum to the intervention effect of Liuwei Dihuang Pill (, LDP) on experimental retinal degeneration, and initially investigate the mechanism of Borneolum syntheticum as meridian-lead-in drug. METHODS: A total of 180 sodium iodateinduced retinital degeneration rats were randomly divided into three groups, including distilled water group, LDP group, and LDP+Borneolum syntheticum (LDP+BS) group. Twenty normal rats were fed regularly without any treatment as normal control. On day 7 and 14 after treatment, histopathological study and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) test were performed to evaluate the retinopathy. Claudin-5 expression at blood-retina barrier (BRB) was detected by Western blot at different time points from 0.5 to 8 h after gavage. RESULTS: On day 7 and 14 after treatment, the retinal lesion grades were significantly different among the three groups (P<0.05). The grade in the LDP+BS group was significantly less than the LDP and distilled water groups (both P<0.05), no significant difference was observed between the LDP and distilled water groups (P>0.05). The apoptosis rates in the LDP+BS group was significantly less than the distilled water and LDP groups (both P<0.05), while there was no significant difference between LDP and distilled water groups (P>0.05). Expression of claudin-5 in LDP+BS group was significantly less than the other two groups at 0.5, 1 and 2 h after gavage (P<0.05). There was no apparent difference among the three groups at 4 and 8 h after gavage (P>0.05). CONCLUSION: Borneolum syntheticum could strengthen the effect of LDP on experimental retinal degeneration, indicated that Borneolum syntheticum might play the role of meridian-lead-in drug in the formula. The mechanism may be due to Borneolum syntheticum could promote the physiologically openness of bloodretina barrier through transiently affecting the expression of claudin-5.

High-resolution melting assay (HRMA) is a simple and sensitive stool-based DNA Test for the detection of mutations in colorectal neoplasms.

BACKGROUND: Stool-based DNA testing for colorectal cancer is becoming a favored alternative to existing DNA screening tests. However, current methods of analysis often become more complicated and costly with increased sensitivity. The high-resolution melting assay (HRMA) is a simple and rapid mutation scanning method with low cost and superb accuracy. In this study, we verified the accuracy of HRMA for screening KRAS/TP53 mutations in stool-isolated DNA from patients with colorectal cancer. MATERIALS AND METHODS: Comparing to direct DNA sequencing, the accuracy of HRMA was verified by detecting KRAS/TP53 mutations in 2 independent stages. In study stage I, both tissue and stool samples from colorectal neoplasm patients were analyzed. In study stage II, stool samples from patients with colorectal neoplasms, and normal controls in clinical screening settings were examined. RESULTS: In study stage I, the HRMA identified 14 of 17 target mutations (82.4%) in stools from cancer patients, and 4 of 5 (80.0%) target mutations in stools from advanced adenoma patients. The mutation detection rate in fecal samples (45.0%; 18/40) and referred tissue samples (55.0%; 22/40) was highly consistent (κ = 0.79). The HRMA detected 1% mutant DNA in a background of wild type DNA. In study stage II, the HRMA assay detected 58.8% (20/34) mutations in tumor samples, 41.5% (17/41) in advanced adenomas samples, and 3.33% (2/60) in age-matched normal control samples. The results from HRMA and DNA sequencing revealed 100% sensitivity and specificity in both tissue and stool samples. CONCLUSION: HRMA is a simple, reliable, and sensitive method for detecting DNA mutations in the stool samples from patients with colorectal neoplasms.