Cell HE staining smears and paired cell paraffin sections in detection of epithelial growth factor receptor gene of pleural fluid specimens / 中南大学学报(医学版)

OBJECTIVES@#The advanced non-small cell lung cancer (NSCLC) patients with pleural effusion have no opportunity for surgery treatment. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the first-line drugs for these patients with EGFR-sensitive mutation. However, the disease progression and drug update during or after treatment of EGFR-TKIs bring more challenges and puzzles to clinical diagnosis and treatment, which inevitably requires archived pleural cell samples for EGFR re-examination or comparative study. Understanding the DNA quality of archived pleural fluid samples and effectively using archival data of pleural fluid cells are of great significance for tracing the origin of cases and basic medical research. This study aims to evaluate the consistency of EGFR mutant gene expression between the 2 methods, and to explore a reliable way for preserving cytological data and making full use of cytological archival data via cell HE staining smear and cell paraffin section.@*METHODS@#A total of 57 pleural fluid cytology cases in the Department of Pathology of China Aerospace Center Hospital from October 2014 to April 2021 were selected. Tumor cells were detected by cell HE staining smears and immunohistochemical staining for TTF-1 and Napsin A in the paired cell paraffin sections. There were more than 200 tumor cells in cell HE staining smear and the proportion of tumor cells were ≥70% in matched cell paraffin sections. Patients with 2 cell smears (one for cell data retention and the other for DNA extraction) were selected as the research subjects, and 57 pleural fluid samples were enrolled. EGFR gene mutation was detected by amplification refractory mutation system-polymerase chain reaction in 57 paired cell HE staining smears and cell paraffin sections. DNA concentration was 2 ng/μL. Cell HE smear was amplified side-by-side with DNA samples from paired cell paraffin sections. Result determination was according to the requirements of the reagent instructions. The external control cycle threshold (Ct) value of the No. 8 well of the samples to be tested was between 13 and 21, which was considered as successful and reliable samples. When the Ct value of EGFR gene mutation was <26, it was considered as positive; when the Ct value was between 26 and 29, it was critical positive; when the Ct value was equal or more than 29, it was negative. ΔCt value was the difference between mutant Ct value and externally controlled Ct value. The smaller the ΔCt value was, the better the quality of DNA of the detected sample was.@*RESULTS@#Among the 57 pleural effusion samples, 42 patients were hospitalized with pleural effusion as the first symptom, accounting for 73.7% (42/57). EGFR mutation was detected in 37 samples [64.9% (37/57)]. The mutation rate for 19del was 37.8% (14/37) while for L858R was 48.6% (18/37). Females were 56.7% (21/37) of mutation cases. The mutation consistency rate of cell HE staining smear and matched cell paraffin sections was 100%. The ΔCt values of cell HE staining smears were less than those of matched cell paraffin sections. The mutation Ct values of 37 cytological samples were statistically analyzed according to the preservation periods of the years of 2014-2015, 2016-2017, 2018-2019, and 2020-2021. There were significant differences in cell paraffin section in the years of 2014-2015 and 2016-2017 compared with the years of 2018-2019 and 2020-2021, while no significant differences were found in cell HE staining smear. Statistical analysis of externally controlled Ct values of 57 cytological samples showed that there were significant differences between cell HE staining smears and cell paraffin section in the years of 2014-2015 and 2016-2017, compared with the years of 2018-2019 and 2020-2021. The mutational Ct values of 37 paired cell blocks and smears were all <26, and the externally controlled Ct values of 57 paired cell paraffin sections and HE staining smears were all between 13 and 21.@*CONCLUSIONS@#The DNA quality of cell HE smears and matched cell paraffin section met the qualified requirements. Two methods possess show an excellent consistency in detecting EGFR mutation in NSCLC pleural fluid samples. The DNA quality of cell HE staining smear is better than that of cell paraffin sections, so cell HE staining smear can be used as important supplement of the gene test source. It should be noted that the limitation of cell HE staining smears is non-reproducibility, so multiple smears of pleural fluid are recommended to be prepared for multiple tests.

Expression of farnesoid X receptor in colorectal cancer and its clinical significance / 肿瘤研究与临床

Objective:To investigate the expression changes of farnesoid X receptor (FXR) in the evolution of normal intestinal mucosa, colorectal adenoma (CRA) and colorectal cancer (CRC), and the correlation of FXR expression with clinicopathological features and prognosis of patients with colorectal tumors.Methods:The UALCAN website tool was used to analyze the expression level of FXR gene transcripts of CRC and normal colorectal tissues in The Cancer Genome Atlas (TCGA) database. The patients undergoing colonoscopy and treatment in the Aerospace Center Hospital from January 2019 to September 2020 were selected, and the immunohistochemistry was used to detect the expression of FXR protein in 100 CRA tissues, 47 CRC tissues and 11 normal colonic mucosal tissues from healthy people (healthy control). Combining with clinical data, the relationship between FXR protein expression and clinicopathological characteristics of patients with colorectal tumors was analyzed. According to the Kaplan-Meier Plotter online database, the median expression level of FXR gene transcripts in CRC patients was analyzed, and the patients were divided into FXR low-expression group and high-expression group, the relationship between the expression of FXR gene and prognosis of CRC patients was investigated.Results:The analysis of data from TCGA database showed that the expression level of FXR gene transcripts in CRC tissues was lower than that in normal colorectal tissues ( P < 0.01). Immunohistochemical examination of the collected tissues showed that the positive rate of FXR protein gradually decreased from the cecum to the rectum. The positive rates of FXR protein in healthy control, CRA patients and CRC patients were 90.9% (10/11), 24.0% (24/100), 6.3% (3/47), and the difference was statistically significant ( χ2 = 35.56, P < 0.01); the positive rate of FXR protein in cancer tissues from CRC patients was lower than that in normal tissues adjacent to cancer [6.3% (3/47) vs. 65.2% (15/23)], and the difference was statistically significant ( χ2 = 27.98, P < 0.01). There was no statistical difference in the positive rate of FXR among CRA patients with different gender, age, maximum diameter of adenoma, and aggression (all P > 0.05). There was also no statistical difference in the positive rate of FXR among CRC patients with different gender, age, tumor site, maximum diameter of tumor, degree of differentiation, TNM staging, and vascular tumor thrombus (all P > 0.05). According to the survival analysis of Kaplan-Meier Plotter online database, the recurrence-free survival of CRC patients with high expression of FXR was better than that of patients with low expression of FXR ( P = 0.003). Conclusions:The expression level of FXR gradually decreases in the intestinal tissues of healthy people, CRA patients and CRC patients. The prognosis of CRC patients with low FXR expression is poor.

Prognostic value of clinical and pathological characteristics in 48 women with pseudomyxoma peritonei / 中华妇产科杂志

Objective To investigate the clinic-pathological characteristics and prognosis of 48 female cases with peritoneal pseudomyxoma(PMP).Methods The clinicopathologic features and follow-up data of 48 female patients with PMP diagnosed in the General Hospital of People's Liberation Army from Jan.1982 to Dec.2011 were retrospectively reviewed.The relationship between clinic-pathological characteristics and prognosis were analyzed using log-rank test and Cox proportional hazards model.Results (1) Clinicopathologic features:the mean age of the 48 cases was 58.8 years (range from 24 to 79 years).Symptoms:abdominal distention and abdominal discomfort were the main symptoms.Imaging examinations showed nonspecific abdominal and pelvic lesions in most cases.Treatment:all the 48 patients underwentlaparotomy and cytoreductive surgery (CRS),in which 15 (31％) patients with completeness of the cancer resection (CCR)-1,24(50％) cases with CCR-2,and CCR-3 in 9(19％) cases.Six (12％) cases were treated by intraoperative hyperthermic intraperitoneal chemotherapy (HIPEC) with cisplatin,20 (42％) patients were treated with different options postoperative adjuvant chemotherapy.Pathological types:the cases were histologically classified into 3 subcategories:disseminated peritoneal adenomucinosis (DPAM),peritoneal mucinous carcinomatosis (PMCA),and PMCA with intermediate or discordant features (PMCA-I/D),which were 22 (46％) cases,9 (19％) cases and 17 (35％) cases,respectively.Appendiceal tumors:44(92％) cases underwent appendectomy,in which 38 cases presented appendiceal tumors (including 20 cases of low-grade appendiceal mucinous adenoma and 18 cases of appendiceal mucinous adenocarcinoma),2 cases were diagnosed as appendicitis,4 cases with unknown pathologic diagnosis.And the other 4(8％) cases,who didn't undergo appendectomy at the first operation,presented peritoneal tumor recurrence and appendiceal mucinous tumors 1,11,32 and 85 months after surgery,respectively.Parenchymal organs involved:ovarian involving was happened in 34 (71％)patients including 15 cases with the right ovary involving,13 cases in both sides,and 6 cases involving the left side.The other parenchymal organs in 10(21％) cases.(2) Prognostic factors:11 patients died,31 survived and 6 cases were lost to follow-up.The mean survival time was 99 months(ranged from 1 to 312 months).The 3-year,5-year and 10-year survival rates were 73.3％,68.0％ and 46.6％,respectively.Univariate statistical analysis showed that age,pathological type and parenchymal involvement were significantly relationship with the survival time (all P ＜ 0.05).But the operation times,appendiceal tumor type,ovarian involvement,CCR,intraperitoneal HIPEC and post-operative adjuvant chemotherapy were not significantly correlate with survival time (all P ＞ 0.05).Multivariate analysis showed that age and pathologic type were independent prognostic factors (P ＜ 0.05).Conclusions No specific clinical features presented in PMP.CRS with HIPEC should the recommended treatment.Both ovaries exploration and appendectomy should be carried out routinely in CRS.The 10-year overall survival of PMP is low.Age,pathological type and parenchymal organs involvement other than ovarian are correlated with the prognosis.And the pathological type and age are independent prognostic factors of PMP.

Preliminary study on grading of injury from adverse drug reaction / 上海交通大学学报（医学版）

Objective To design a quantitative criterion and grading system for injury from adverse drug reaction (ADR) in order to serve the compensation system of injury from ADR. Methods Based on the other grading system of injury cases, the independent scores and serial grades were given to the ADR injury of different organs or different levels through two turns of consultation to 27 experts from Shanghai. Results Injury from ADR was graded from stage 1 to stage 10 according to the damage degree, with 1 stand for death and 10 for slightest injury. Conclusion The grading method of ADR based on the characteristics of injury accords with the condition of China, and can provide reference for making up the compensation system of ADR injury.