ABSTRACT
BACKGROUND: HER2 status assessment is an Important biological Index for the treatment and prognosis of Invasive breast cancer. Pre-treatment of tissues such as Immobilization, dehydration, transparency and dewaxlng Is a necessary procedure for HER2 protein and gene detection after pathological paraffin sections, and also an important factor affecting Immunohlstochemlstry and fluorescence in situ hybridization. OBJECTIVE: To explore the application value of environment-friendly biological tissue sample preparation kit In fluorescence in situ hybridization detection of HER2 protein 2-positive Invasive breast cancer. METHODS: 402 Invasive breast cancer specimens were collected from Shantou Central Hospital from January 2015 to March 2019. The same specimens were semi-dissected and randomly divided Into two groups. The control group was treated by traditional reagent formaldehyde immobilization-ethanol dehydratlon-xylene transparency and dewaxlng, and paraffin sections were made. The experimental group was treated with formaldehyde immobilization-ethanol dehydratlon-xylene transparent dewaxlng. Environment-friendly biological tissue sample preparation kit (Including environmentally friendly stationary fluid, dehydration fluid, transparent liquid, dewaxing fluid) was used to make slices. The expression of HER2 protein was detected by Immunohlstochemlstry. The amplification of HER2 gene was detected by fluorescence in situ hybridization In 131 Invasive breast cancer specimens with positive HER2 protein expression. RESULTS AND CONCLUSION: The expression of HER2 protein In both experimental and control groups was specific and cell localization was correct. There were no significant differences In HER2 protein positive rate, uncertainty rate, and negative rate between the two groups (P > 0.05).The coincidence rate of HER2 protein expression between the two groups was 99.00%. The background of HER2 gene was clear In both groups, and the signals of HER2 and CM 7 double probes were clear. There was no cross-reaction and the double probe signal was precisely located In the nucleus of cancer cells. There was no significant difference In the number of successful cells between the two groups (P > 0.05). There was no significant difference in the positive rate and negative rate of HER2 gene amplification between the two groups (P > 0.05). The coincidence rate of HER2 gene amplification between the two groups was 97.71%. The average signal number of HER2 gene and the ratio of HER2/cells In both groups were all equal. There was no significant difference in the mean number of Ch17 signal, Ch17/cell ratio and HER2/CM7 ratio between the two groups (P > 0.05). There was no significant difference in the total positive rate of HER2 between the two groups (P > 0.05). The results showed that compared with the traditional reagents, the invasive breast cancer samples prepared by environment-friendly bio-tissue sample preparation kit had no effect on HER2 protein expression. The expression of HER2 protein does not affect the amplification of HER2 gene, which can meet the needs of clinical detection.
ABSTRACT
BACKGROUND: The free radicals induced by cerebral ischemia/reperfusion consist mainly of xanthine oxidase, which induces cell swelling in the infarcted area.OBJECTIVE: To observe the changes of cerebral ischemia/reperfusioninduced changes in the activity of cerebral superoxide dismutase (SOD), an enzyme responsible for free radical clearance, and investigate the effect of apurin, a inhibitor of purine oxidase, on cellular water content in the brain tissue with ischemia/reperfusion injury.DESIGN: Completely randomized controlled study.SETTING: Department of Neurology of the Central Hospital Affiliated to Shenyang Medical College, Department of Neurosurgery of the First Hospital Affiliated to China Medical University, and Liaoning Provincial Orthopedic Hospital for Limb Disabilities.MATERIALS: The experiment was conducted in the Laboratory of the Central Hospital Affiliated to Shenyang Medical College from May 2003 to April 2004. Forty Wistar rats were subjected to a 6-hour cerebral ischemia and randomized into 4 equal groups to receive intragastric administration of 100 mg/kg apurin (ischemia + apurin group), oxolinic acid suspension of the same dose (ischemia+ oxolinic acid group), 100 mg/kg apurin after a 2-hour reperfusion (Ischemia/reperfusion + apurin group), or oxolinic acid of the same dosage after the 2-hour reperfusion (ischemia/reperfusion + oxolinic acid goup), respectively. The rats in apurin group had intragastric administration of 100 mg/kg apurin 48, 24 and 1 hour before occlusion of the cervical internal carotid artery (CICA) to induce the ischemia, respectively. Oxolinic acid was given in the two oxolinic acid groups in the same manner.METHODS:Water content of brain tissue of rats was measured after 6 hours of CICA occlusion in the two ischemia groups and after the 2-hour perfusion in the two ischemia/reperfusion groups. Distribution of SOD in the brain tissue was observed with SOD immunostaining.MAIN OUTCOME MEASURES: Distribution of SOD and water content in the brain tissue of rats.RESULTS: In the two oxolinic acid groups, Cu-Zn SOD staining identified obviously increased staining intensity in the ischemic foci. Mn SOD staining in ischemia+oxolinic acid group resulted in increased circular staining surrounding the vessels in the ischemic foci, with also obvious staining of the vascular wall and neural cells. The ischemic foci of the ischemia/reperfusion + oxolinic acid group showed diffuse but lightly weaker staining. Cu-Zn SOD staining in the two apurin groups revealed no significant difference. In the two oxolinic acid groups, endothelial cell nuclear swelling of the arteriole, protrusion of the mid-layer myocytes, and expansion of the vascular membrane were observed, with the tissues surrounding the vessels appearing spongy. These changes were less severe in the two apurin groups. The water content in the brain tissue was (78.56±0.30) % in ischemia + apurin group and (78.85±0.49) % in ischemia/reperfusion + apurin group, significantly lower than that of (79.08±0.33) % in ischemia + oxolinic acid group and (79.86±0.49) % in ischemia/reperfusion + oxolinic acid group (P ＜ 0.05).CONCLUSION: Apurin can relieve tissue injury after cerebral ischemia/reperfusion by inhibition of SOD.