Human Malignant Rhabdoid Tumor Antigens as Biomarkers and Potential Therapeutic Targets.

INTRODUCTION: Atypical teratoid rhabdoid tumor (ATRT) is a lethal type of malignant rhabdoid tumor in the brain, seen mostly in children under two years old. ATRT is mainly linked to the biallelic inactivation of the SMARCB1 gene. To understand the deadly characteristics of ATRT and develop novel diagnostic and immunotherapy strategies for the treatment of ATRT, this study investigated tumor antigens, such as alpha-fetoprotein (AFP), mucin-16 (MUC16/CA125), and osteopontin (OPN), and extracellular matrix modulators, such as matrix metalloproteinases (MMPs), in different human malignant rhabdoid tumor cell lines. In addition, the roles of MMPs were also examined. MATERIALS AND METHODS: Five human cell lines were chosen for this study, including two ATRT cell lines, CHLA-02-ATRT and CHLA-05-ATRT; a kidney malignant rhabdoid tumor cell line, G401; and two control cell lines, human embryonic kidney HEK293 and HEK293T. Both ATRT cell lines were treated with a broad-spectrum MMP inhibitor, GM6001, to investigate the effect of MMPs on cell proliferation, viability, and expression of tumor antigens and biomarkers. Gene expression was examined using a reverse transcription polymerase chain reaction (RT-PCR), and protein expression was characterized by immunocytochemistry and flow cytometry. RESULTS: All the rhabdoid tumor cell lines tested had high gene expression levels of MUC16, OPN, AFP, and MSLN. Low expression levels of neuron-specific enolase (ENO2) by the two ATRT cell lines demonstrated their lack of neuronal genotype. Membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14) and tissue inhibitor of metalloproteinases-2 (TIMP-2) were highly expressed in these malignant rhabdoid tumor cells, indicating their invasive phenotypes. GM6001 significantly decreased ATRT cell proliferation and the gene expression of MSLN, OPN, and several mesenchymal markers, suggesting that inhibition of MMPs may reduce the aggressiveness of rhabdoid cancer cells. CONCLUSION: The results obtained from this study may advance our knowledge of the molecular landscapes of human malignant rhabdoid tumors and their biomarkers for effective diagnosis and treatment. This work analyzed the expression of human malignant rhabdoid tumor antigens that may serve as biomarkers for the development of novel therapeutic strategies, such as cancer vaccines and targeted and immunotherapies targeting osteopontin and mesothelin, for the treatment of patients with ATRT and other malignant rhabdoid tumors.

Duodenal cholinergic tuft cell number is increased in functional dyspepsia.

BACKGROUND: Low-grade duodenal inflammation has recently been identified in patients with functional dyspepsia (FD). Chemosensory tuft cells were reported to be associated with gastrointestinal diseases. We therefore assessed duodenal tuft cell density and microinflammation in patients with FD to determine whether these measures could serve as useful biomarkers, and also correlated tuft cell density and microinflammation in FD patients. METHODS: Duodenal biopsy specimens were obtained from patients with FD and from controls. Tuft cells, eosinophils, and mast cells were immunochemically stained with specific antibodies. Tuft cells were identified by immunostaining for doublecortin-like kinase 1 (DCLK1); cholinergic tuft cells were assessed by double staining for choline acetyltransferase (ChAT) and DCLK1. Immune-type tuft cells were assessed by IL-25 mRNA expression using real-time PCR. KEY RESULTS: The density of intramucosal eosinophils and mast cells was significantly higher in the duodenum of FD patients than in controls. The density of tuft cells was significantly higher in the duodenum of FD patients compared with controls, and significantly correlated with eosinophil density in the duodenum of FD patients and controls. Moreover, a fraction of ChAT-positive cells was DCLK1 positive; all duodenal DCLK1+ tuft cells were ChAT-immunoreactive in FD and in control subjects. CONCLUSIONS AND INFERENCES: Cholinergic tuft cell density was higher in the duodenum of patients with FD and significantly correlated with eosinophil density. Further studies are needed to investigate the pathophysiological significance of tuft cells in FD and may provide valuable clues to the pathophysiology of FD.

Metagenomic Assessment of the Pathogenic Risk of Microorganisms in Sputum of Postoperative Patients With Pulmonary Infection.

Respiratory infections are complicated biological processes associated with an unbalanced microbial community and a wide range of pathogens. To date, robust approaches are still required for distinguishing the pathogenic microorganisms from the colonizing ones in the clinical specimens with complex infection. In this study, we retrospectively analyzed the data of conventional culture testing and metagenomic next-generation sequencing (mNGS) of the sputum samples collected from 50 pulmonary infected patients after cardiac surgery from December 2020 and June 2021 in Ruijin Hospital. Taxonomic classification of the sputum metagenomes showed that the numbers of species belonging to bacteria, fungi, and viruses were 682, 58, and 21, respectively. The full spectrum of microorganisms present in the sputum microbiome covered all the species identified by culture, including 12 bacterial species and two fungal species. Based on species-level microbiome profiling, a reference catalog of microbial abundance detection limits was constructed to assess the pathogenic risks of individual microorganisms in the specimens. The proposed screening procedure detected 64 bacterial pathogens, 10 fungal pathogens, and three viruses. In particular, certain opportunistic pathogenic strains can be distinguished from the colonizing ones in the individual specimens. Strain-level identification and phylogenetic analysis were further performed to decipher molecular epidemiological characteristics of four opportunistic etiologic agents, including Klebsiella pneumoniae, Corynebacterium striatum, Staphylococcus aureus, and Candida albicans. Our findings provide a novel metagenomic insight into precision diagnosis for clinically relevant microbes, especially for opportunistic pathogens in the clinical setting.

Three-year outcomes of transanal total mesorectal excision versus standard laparoscopic total mesorectal excision for mid and low rectal cancer.

INTRODUCTION: Since transanal total mesorectal excision (taTME) was introduced, it has become an important topic in rectal cancer treatment. Many previous studies reported positive relevant short-term results, histopathological results, and associated complications. Recently, concerns regarding the oncological safety of taTME have been raised due to reports showing high local recurrences (LR) rates. Therefore, this study aimed to compare the 3-year outcomes between taTME and laparoscopic total mesorectal excision (laTME) for mid-low rectal cancer. METHODS: A total of 104 patients who underwent taTME were matched with 208 patients treated by laTME. The primary endpoint was 3-year LR rate; secondary endpoints in this matched-cohort study included the perioperative outcomes and histopathological outcomes. RESULTS: taTME was associated with lower permanent ostomy rate (1% vs 13.5%) and lower conversion rate (0% vs 3.4%) compared to laTME. A similar quality of resected specimens was detected for each group. In both groups, the local recurrence rate was 3.8%. Within 3 years after surgery, the disease-free survival (DFS) rates were 78.8% in the taTME group and 76.9% in the laTME group (P = 0.640), while the overall survival (OS) rates were 93.3% in the taTME group and 89.9% in the laTME group (P = 0.327). CONCLUSION: No significant differences regarding 3-year local recurrence rate (3.8%) were observed in the taTME group compared to laTME group.

Photothermal therapy technology of metastatic colorectal cancer.

Colorectal cancer (CRC) is one of the most common malignancies. The current treatments of metastatic colorectal cancer (mCRC) are ineffective and the bottleneck problem. It is of significance to explore effective new therapeutic strategies to eradicate mCRC. Photothermal therapy (PTT) is an emerging technology for tumor therapy, with the potential in the treatment of mCRC. In this review, the current treatment approaches to mCRC including surgery, radiotherapy, chemotherapy interventional therapy, biotherapy, and photothermal therapy are reviewed. In addition, we will focus on the various kinds of nanomaterials used in PTT for the treatment of CRC both in vitro and in vivo models. In conclusion, we will summarize the combined application of PTT with other theranostic methods, and propose future research directions of PTT in the treatment of CRC.

Targeting and imaging colorectal cancer by activatable cell-penetrating peptides.

While it has been a great challenge to determine the positive status of metastasis lesions, intraoperative tumor imaging, which can show tumor localization and facilitate intraoperative staging of nodal metastases, have enabled surgeons to quickly and accurately perform radical resections. However, to date, there is no accurate method for evaluating nodal status intraoperatively. In this study, we synthesized activatable cell-penetrating peptides (ACPPs) that can specifically recognize colorectal cancer and their nodal status. ACPPs were labeled with Cy5 dye at the C-terminal, and named ACPP-Cy5. Laser scanning confocal microscopy and flow cytometry were used to measure the change in intracellular fluorescence intensity between cancer cells and normal cells. The results showed while the intracellular Cy5 fluorescent intensity can be visualized in both cancer and normal cells by 8 h after adding ACPP-Cy5, the relative fluorescence intensity of colorectal cancer cells was significantly higher than the normal cells. In addition, IVIS spectrum in vivo imaging system was used to observe the fluorescence intensity of ACPP-Cy5 after tail vein injection of mice with subcutaneous tumor or orthotopic colorectal cancer and liver metastasis. We found in mice with colorectal cancer and liver metastasis the Cy5 fluorescence intensity of cancer was significantly increased compared to the organs including liver, colorectum, lung, spleen, and heart. It is demonstrated here, this ACPPs can target colorectal cancer and liver metastasis, therefore ACPP-Cy5 may be a promising tool used for the diagnoses of colorectal cancer and to assist in tumor localization during surgery.

Comparison of pathological outcomes after transanal versus laparoscopic total mesorectal excision: a prospective study using data from randomized control trial.

INTRODUCTION: Total mesorectal excision (TME) is the standard procedure for middle lower rectal cancer, and transanal total mesorectal excision (taTME) was founded as a valid alternative to the open and laparoscopic TME. The quality of the procedure performed is important for prognosis of patients. This study was designed to compare the pathological results of taTME with those of laparoscopic TME (laTME), based on the data from a randomized control trial (RCT: NCT02966483). METHODS: Between April 2016 and November 2018, all rectal cancer patients who underwent taTME or laTME in the Sixth Affiliated Hospital of Sun Yat-sen University (Guangzhou, China) and enrolled in the RCT were included in this study. The data from all participants were prospectively input in a standardized database. RESULTS: In total 128 patients were included in the taTME group and 133 patients were included in the laTME group. The demographics and tumor characteristics were not significantly different between the two group. T3 or N0 lesions were most common in both groups. The mesorectum specimen was complete or nearly complete in all patients. The positive distal resection margin (DRM) was detected in 2 (1.5%) cases in the laTME group versus no cases in the taTME group (P = 0.498), and the distance between the tumor and DRM in the taTME group (1.4 ± 1.1) may have the longer tendency than that in the laTME group (1.3 ± 0.9) (P = 0.745). The positive circumferential resection margin was detected in 2 cases in each group (P = 0.674). The median number of resected lymph nodes was 15.0 in taTME group versus 16.0 in the laTME group (P = 0.069). CONCLUSION: The pathological outcomes between transanal and laparoscopic total mesorectal excision are similar. The rate of positive resection margin could not be significant decreased, nonetheless the decrease trend could be shown.

Safety and Feasibility of Transanal Endoscopic Surgery for Diffuse Cavernous Hemangioma of the Rectum.

PURPOSE: To evaluate the safety and feasibility of transanal endoscopic surgery for diffuse cavernous hemangioma of the rectum (DCHR). METHODS: All DCHR patients who underwent transanal endoscopic surgery in our hospital between January 2014 and June 2018 were reviewed. RESULTS: A total of 7 patients with a diagnosis of DCHR underwent transanal endoscopic surgery during the study period. Four patients (57.1%) were male, with a mean age at surgery of 34.5 ± 7.7 years, and three patients (42.9%) were female, with a mean age at surgery of 29.9 ± 3.8 years. Recurrent painless rectal bleeding was the main symptom in all patients. The mean age was 32 years old (range 21-54 years). The median duration of symptoms was 10 years (range 1 month-50 years). The level of hemoglobin at admission ranged from 59.0 to 148.0 g/l (mean 106.6 g/l), and the level of mean corpuscular volume (MCV) ranged from 75.1 fl to 93.5 fl (mean 83.7 fl). Colonoscopy, computed tomography (CT), and magnetic resonance imaging (MRI) were important in the diagnosis of DCHR because of their high positive rates and accurate features. All of the lesions are between the anal canal and the descending colon. Two patients could be found with some enlarged serpentine vessels in the cervix, vagina, or corpus cavernosum by MRI. After admission, all the patients underwent transanal endoscopic surgery and four patients had simultaneous loop ileostomy. The mean operative time was 278 min (range 168-400 min). The median amount of intraoperative blood loss was 50 ml (range 10-300 ml). The mean distance from anal verge to anastomosis was 2.2 ± 0.2 cm. The anastomosis was fashioned with a stapler in two patients (28.6%). There were no intraoperative and postoperative complications. All the patients continued to recover well from the surgery, and nobody needed postoperative blood transfusions. CONCLUSIONS: The specific diagnosis rate of DCHR is low. Preoperative MRI and CT examination can make a definitive diagnosis and determine the extent of the lesions. DCHR is mostly restricted to the rectum, sigmoid colon, anal wall, and mesorectum. The best treatment for DCHR is complete lesion resection. It is safe and feasible to treat DCHR using transanal endoscopic surgery. Moreover, transanal endoscopic surgery might have a huge potential when used to treat other rectal diseases.

Prevalence of Fabry disease in stroke patients--a systematic review and meta-analysis.

BACKGROUND: Fabry disease is an uncommon but treatable cause of stroke. Enzyme replacement therapy helps improve neurologic symptoms. We conducted a systematic review and meta-analysis to evaluate the prevalence of Fabry disease in stroke patients. METHODS: We searched MEDLINE and EMBASE databases for relevant articles published in English up to February 2013. Studies that reported incidence or prevalence of Fabry disease in stroke patients were included. Two reviewers independently assessed studies to determine eligibility, validity, and quality. Meta-analysis was performed to calculate the prevalence of Fabry disease by etiology and gender. RESULTS: Nine studies (n = 8302 patients) met the inclusion criteria. Eight studies (n = 8148) examined the prevalence of Fabry disease in young stroke patients. Overall qualities of included studies were moderate to high. The prevalence of Fabry disease ranged from .4% to 2.6% on strokes of any etiologies. In cryptogenic stroke, the prevalence ranged from .6% to 11.1%, 4.5% in men (95% confidence interval [CI] = 3.2%-6.3%) and 3.4% in women (95% CI = 1.0%-10.7%). The prevalence of Fabry disease in patients with all etiologies was similar in men (.9% [95% CI = .3%-2.3%]) and (1.4% [95% CI = .7%-2.7%]) in women. CONCLUSIONS: Fabry disease may explain approximately 1% of all strokes in the young, including 3%-5% of cryptogenic strokes. The confirmation of Fabry disease may be challenging as there are different criteria for men and women. Early recognition of Fabry disease may help initiate the appropriate treatment to decrease the risk of subsequent complications.

CCR4, a 3'-5' poly(A) RNA and ssDNA exonuclease, is the catalytic component of the cytoplasmic deadenylase.

The CCR4-NOT complex from Saccharomyces cerevisiae is a general transcriptional regulatory complex. The proteins of this complex are involved in several aspects of mRNA metabolism, including transcription initiation and elongation and mRNA degradation. The evolutionarily conserved CCR4 protein, which is part of the cytoplasmic deadenylase, contains a C-terminal domain that displays homology to an Mg2+-dependent DNase/phosphatase family of proteins. We have analyzed the putative enzymatic properties of CCR4 and have found that it contains both RNA and single-stranded DNA 3'-5' exonuclease activities. CCR4 displays a preference for RNA and for 3' poly(A) substrates, implicating it as the catalytic component of the cytoplasmic deadenylase. Mutations in the key, conserved catalytic residues in the CCR4 exonuclease domain abolished both its in vitro activities and its in vivo functions. Importantly, CCR4 was active as a monomer and remained active in the absence of CAF1, which links CCR4 to the remainder of the CCR4-NOT complex components. These results establish that CCR4 and most probably other members of a widely distributed CCR4-like family of proteins constitute a novel class of RNA-DNA exonucleases. The various regulatory effects of the CCR4-NOT complex on gene expression may be executed in part through these CCR4 exonuclease activities.