Recent studies have demonstrated that the brain is equipped with a lymphatic drainage system that is actively involved in parenchymal waste clearance, brain homeostasis and immune regulation. However, the exact anatomic drainage routes of brain lymph fluid (BLF) remain elusive, hampering the physiological study and clinical application of this system. In this study, we systematically dissected the anatomy of the BLF pathways in a rat model. Moreover, we developed a protocol to collect BLF from the afferent lymphatic vessels of deep cervical lymph nodes (dcLNs) and cerebrospinal fluid (CSF) from the fourth ventricle. Nuclear magnetic resonance spectroscopy showed that BLF contains more metabolites than CSF, suggesting that BLF might be a more sensitive indicator of brain dynamics under physiological and pathological conditions. Finally, we identified several metabolites as potential diagnostic biomarkers for glioma, Parkinson's disease and CNS infectious diseases. Together, these data may provide insight into the physiology of the lymphatic system in the brain and into the clinical diagnosis of CNS disorders.
Objective
Immunohistochemical (IHC) staining for mismatch repair (MMR) proteins and PCR-based detection for microsatellite status are routinely performed on colorectal carcinoma (CRC) surgical samples. However, the concordance of the two detections, which is related to the quality control of our molecular pathology laboratory, is unknown so far. So the main aim of this study is to compare the differences between the two analyses and to improve our work.
Methods
IHC analyzed the expression of MLH1, PMS2, MSH2 and MSH6 which was performed on 368 cases of formalin-fixed paraffin-embedded (FFPE) CRC tissues. If any one protein is negative in all of cancer cells but positive in normal colorectal mucosa, the IHC staining was reported as mismatch repair defective (dMMR). If the four MMR proteins are expressed in the nucleus of one or more cancer cells, the IHC result was interpreted as mismatch repair proficient (pMMR). All of the 37 cases of dMMR and selected 28 cases of pMMR were tested by PCR-based MSI analysis. Paired normal and cancer DNA samples isolated from the FFPE tissues were tested for MSI using Bethesda recommended 5 markers (BAT25, BAT26, D2S123, D5S346, D17S250). At last the results of IHC and PCR were compared and their concordance were analyzed.
Results
IHC analyses were performed on 368 cases of CRC, among which 37 cases were dMMR and 331 cases were pMMR. After excluding 2 cases from the 37 samples, the remained 35 samples were tested for MSI, among which 32 samples were high-level microsatellite instability (MSI-H) and 3 samples were microsatellite stable (MSS). In addition, 28 cases of pMMR samples were selected to be tested by PCR for MSI, among which 27 cases were MSS but one case was MSI-H. The sensitivity and specificity of immunohistochemistry was 97.0% and 90.0%, separately, the sensitivity and specificity of PCR was 91.4% and 96.4%, separately, and the total concordance of the two detections achieved 93.7%. The most common original site of dMMR CRC was right hemicolon (occupying 48.6%), and the most common pathological features included mucous adenocarcinoma, poor differentiated adenocarcinoma with lymphocytes infiltration, and pathologic TNM stage Ⅱ and stage Ⅲ.
Conclusions
The total concordance of the immunohistochemistry for MMR proteins and PCR-based MSI testing achieved 93.7%, and the former is an economic and quick screening method which is deserved to be popularized in China. Moreover, we have to emphasize the important role of intra and external laboratory quality control of the two methods and then to improve the process, so as to increase the testing accuracy.
Key words:
Colorectal neoplasms; Immunohistochemistry; Microsatellite instability; Mismatch repair proteins
Mucinous cystadenocarcinoma (MCA) of the breast is extremely rare and was only recently described as a distinct variant of invasive ductal carcinoma of the breast. A case of MCA is reported in a 41‐year‐old woman. Mammographic and ultrasonographic examinations showed an irregularly shaped 10.0 × 8.0 × 5.5 cm lesion with patching calcification in the upper outer quadrant of the left breast. The gross examination revealed that the tumor has a well‐circumscribed edge with a gelatinous cut surface and hemorrhage and necrosis were also noticed in the mass. Microscopically, the mass resembled mucinous cystic neoplasm of the ovary and pancreas closely, with cystic areas lined by columnar mucinous cells and associated with abundant extracellular and intracellular mucin, which is distinctively different from mucinous carcinoma with typically nests of low grade neoplastic cells floating in the mucin pool. The tumor cells were positive for CK7, CK20 and CDX2 were negative and displayed a typical immunophenotype of basal‐like breast cancer (ER, PR, HER2 were negative, CK5/6 and EGFR were positive). Metastatic carcinoma was identified in three of 14 axillary lymph nodes. We describe here a very unusual case of breast MCA with basal‐like immunophenotype.
With the increased uses of targeted therapeutics, diagnostic detection of target mutations becomes essential for the effective clinical applications of targeted therapeutics. Currently, there are two types of methods detecting target mutations in clinics: one is based on DNA sequence and the other uses the newly developed mutation-specific antibodies recognizing mutated proteins. Each method has its own advantages and disadvantages. Here, we explored the sensitivity and specificity of a new commercially available BRAF(V600E) mutation-specific mouse monoclonal antibody. Using routine manual immunohistochemistry (IHC), we tested tumor tissues from 38 melanoma patients. For those melanoma tissues with abundant endogenous melanin, we pretreated the tumor tissues with 3 % hydrogen peroxide to remove melanin for reliable signal detection. We also performed DNA sequencing and ARMS-PCR analyses for these 38 tumor samples. Comparing to the results from DNA-based detection methods, the IHC method with this BRAF(V600E) mutation-specific antibody displayed 100 % sensitivity and 92.9 % specificity. Hence, this IHC detection is sensitive for clinic uses as a simple, fast, inexpensive, and reliable method to screen cancer patients for the BRAF(V600E) mutation and could be easily adapted for use in most hospital pathology laboratories.
The WHO recommends moderate physical activity to combat the increasing risk of death from chronic diseases. We conducted a meta-analysis to assess the association between physical activity and cancer mortality and the WHO recommendations to reduce the latter.MEDLINE and EMBASE were searched up until May 2014 for cohort studies examining physical activity and cancer mortality in the general population and cancer survivors. Combined HRs were estimated using fixed-effect or random-effect meta-analysis of binary analysis. Associated HRs with defined increments and recommended levels of recreational physical activity were estimated by two-stage random-effects dose-response meta-analysis.A total of 71 cohort studies met the inclusion criteria and were analysed. Binary analyses determined that individuals who participated in the most physical activity had an HR of 0.83 (95% CI 0.79 to 0.87) and 0.78 (95% CI 0.74 to 0.84) for cancer mortality in the general population and among cancer survivors, respectively. There was an inverse non-linear dose-response between the effects of physical activity and cancer mortality. In the general population, a minimum of 2.5 h/week of moderate-intensity activity led to a significant 13% reduction in cancer mortality. Cancer survivors who completed 15 metabolic equivalents of task (MET)-h/week of physical activity had a 27% lower risk of cancer mortality. A greater protective effect occurred in cancer survivors undertaking physical activity postdiagnosis versus prediagnosis, where 15 MET-h/week decreased the risk by 35% and 21%, respectively.Our meta-analysis supports that current physical activity recommendations from WHO reduce cancer mortality in both the general population and cancer survivors. We infer that physical activity after a cancer diagnosis may result in significant protection among cancer survivors.
Plasmon-enhanced fluorescence (PEF)-based analytical technology has recently demonstrated its ability in detecting biomarkers with ultrahigh sensitivity. However, the scope of the PEF-based technology has been hindered by its reliance on flat substrates with relatively low binding kinetics and the limited multiplex detection ability. Herein, we reported a simple yet robust method for the fabrication of plasmonic magnetic microbeads (PMMBs)-based suspension array technology (SAT) with fluorescence enhancement of about 60-fold, improving the detection limit of biomarkers by 2-orders of magnitude toward 100 fM. We also demonstrated the performance of this method for the detection of anti-acidic ribosomal phosphoprotein 0 (anti-P0) autoantibody in sera from systemic lupus erythematosus (SLE) patients. Owing to the high sensitivity and efficient magnet-based sample collection, our method can be employed for detection of ultrasmall volumes of samples (e.g., 2 μL), promising for point-of-care detection. Furthermore, a size-encoded PMMBs-based multiplexed suspension array for simultaneous detection of multiple biomarkers is realized, illustrating the great potential of this technology in high-throughput disease diagnosis applications.
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