Rectal Tissue Research Articles

Comparison of microbial signatures between paired faecal and rectal biopsy samples from healthy volunteers using next-generation sequencing and culturomics

BackgroundFaecal samples are frequently used to characterise the gut microbiota in health and disease, yet there is considerable debate about how representative faecal bacterial profiles are of the overall gut community. A particular concern is whether bacterial populations associated with the gut mucosa are properly represented in faecal samples, since these communities are considered critical in the aetiology of gastrointestinal diseases. In this study we compared the profiles of the faecal and mucosal microbiota from ten healthy volunteers using bacterial culturing (culturomics) and next-generation sequencing targeting the 16S ribosomal nucleic acid (rRNA) gene. Paired fresh rectal biopsies and faecal samples were processed under stringent anaerobic conditions to maintain the viability of the bacteria. Four different sample types were analysed: faecal (F), faecal homogenised (FHg), biopsy tissue (B) and biopsy wash (BW) samples. ResultsThere were no significant statistical differences in either bacterial richness or diversity between biopsy washes (BW) and faecal (F) or faecal homogenised (FHg) samples. Principal coordinates analysis of a Bray–Curtis distance matrix generated from sequence variant tables did not show distinct clustering between these samples (PERMANOVA; p = 0.972) but showed strong clustering of samples from individual donors. However, the rectal biopsy tissue (B) samples had a significantly altered bacterial signature with greater abundance of Proteobacteria and Acidobacteria compared to faecal (F) and faecal homogenised (FHg) samples. A total of 528 bacteria encompassing 92 distinct bacterial species were isolated and cultured from a subset of six volunteer samples (biopsy washes and faeces). This included isolation of 22 novel bacterial species. There was significant similarity between the bacterial species grown in anaerobic culture and those identified by 16S rRNA gene sequencing (Spearman correlation; rho = 0.548, p = 0.001).ConclusionThis study showed that the bacterial profiles of paired faecal and rectal biopsy wash samples were very similar, validating the use of faecal samples as a convenient surrogate for rectal biopsy-associated microbiota. Anaerobic bacterial culture results showed similar taxonomic patterns to the amplicon sequence analysis disproving the dogma that culture-based methods do not reflect findings of molecular assessments of gut bacterial composition.FGviAy1dgRvRfr2cUx9EBxVideo abstract

The complementary role of lymphovascular invasion and perineural invasion in the TNM staging process of rectal cancer.

The aim of this study is to clarify the association between lymphovascular invasion (LVI) and/or perineural invasion (PNI) and the clinical characteristics and prognostic importance of rectal cancer, to provide a basis for early adjuvant treatment of rectal cancer. We retrospectively analyzed patients diagnosed with rectal cancer. This study involved rectal cancer tissue samples were obtained by surgical methods. Data on histological form, tumor classification, tumor size, gross growth pattern, blood and lymphatic vessel invasion, and PNI of the slice by HE staining were obtained from pathological examination. Immunohistochemical analysis of tissue samples was performed to determine p53 and EGFR expressions. There were 330 rectal cancer patients included in the study. LVI and/or PNI can be used as a high-risk factor for the prognosis of rectal cancer, predict prognostic survival, and guide adjuvant therapy. The detection rates of LVI and PNI were 32.1% and 16.1%. Differentiation grade, Union for International Cancer Control staging, tumor-lymph node-metastasis staging are significantly related to LVI or PNI. Multivariate logistic regression analysis shows that poor differentiation and N ≥ 1 can be used as independent risk factors and predictive factors for LVI. At the same time, poor differentiation and T > 3 is an independent risk factor for PNI. Only poor differentiation is the risk factor for poor prognosis in Cox risk regression analysis. In addition, the simultaneous occurrence of LVI and PNI is an independent prognostic factor.

Expression of PD-L1 and PD-1 in stage T4 rectal cancer tissues and surrounding metastatic lymph nodes and correlation with prognosis.

The expressions of programmed death ligand 1 (PD-L1) and programmed death receptor 1 (PD-1) in T4 rectal cancer tissues and surrounding metastatic lymph nodes were analyzed and correlated with prognosis. For this purpose, ninety-eight patients with T4 rectal cancer treated in our hospital from July 2021 to July 2022 were selected, and surgically resected rectal cancer tissues as well as para carcinoma tissue samples, and surrounding metastatic lymph node tissues were obtained from all patients. Analysis of PD-L1 and PD-1 expression in rectal cancer tissues as well as in adjacent tissue specimens and surrounding metastatic lymph node tissues were performed using immunohistochemical staining. PD-L1 and PD-1 expression were analyzed in relation to lymph node metastasis, maximum tumor diameter, as well as histological analysis, and the relationship between the two and prognosis was analyzed. Immunohistochemistry for PD-L1, PD-1 revealed that both proteins were expressed in association with the target cytoplasm as well as within the cell membrane; The number of cases with positive expression of PD-L1 and PD-1 in cancer tissues was significantly higher than the number of cases with expression in adjacent tissues, which was statistically significant (P < 0.05); The expression rates of PD-L1, PD-1 in poor expression in progression-free survival as well as in progression survival were significantly higher than those in medium and high expression with statistical significance (P < 0.05); Compared with patients without lymph node metastasis, patients with T4 rectal cancer with lymph node metastasis had a higher number of cases with high expression levels of PD-L1 and PD-1 proteins, and the difference was statistically significant (P < 0.05); PD-L1 and PD-1 in T4 stage rectal cancer prognosis is closely related, distant metastasis as well as lymph node metastasis has a greater effect on PD-L1 and PD-1. PD-L1 and PD-1 showed abnormal expression in T4 rectal cancer tissues as well as in surrounding metastatic lymph nodes, and PD-L1 and PD-1 were closely related to prognosis in T4 rectal cancer, distant metastasis, as well as lymph node metastasis had a greater effect on PD-L1 and PD-1. Its detection is able to provide a certain data reference for the prognosis of T4 rectal cancer.

Presume Why Probiotics May Not Provide Protection in Inflammatory Bowel Disease through an Azoxymethane and Dextran Sodium Sulfate Murine Model.

Recent studies have shown dysbiosis is associated with inflammatory bowel disease (IBD). However, trying to restore microbial diversity via fecal microbiota transplantation (FMT) or probiotic intervention fails to achieve clinical benefit in IBD patients. We performed a probiotic intervention on a simulated IBD murine model to clarify their relationship. IBD was simulated by the protocol of azoxymethane and dextran sodium sulfate (AOM/DSS) to set up a colitis and colitis-associated neoplasm model on BALB/c mice. A single probiotic intervention using Clostridium butyricum Miyairi (CBM) on AOM/DSS mice to clarify the role of probiotic in colitis, colitis-associated neoplasm, gut microbiota, and immune cytokines was performed. We found dysbiosis occurred in AOM/DSS mice. The CBM intervention on AOM/DSS mice failed to improve colitis and colitis-associated neoplasms but changed microbial composition and unexpectedly increased expression of proinflammatory IL-17A in rectal tissue. We hypothesized that the probiotic intervention caused dysbiosis. To clarify the result, we performed inverse FMT using feces from AOM/DSS mice to normal recipients to validate the pathogenic effect of dysbiosis from AOM/DSS mice and found mice on inverse FMT did develop colitis and colon neoplasms. We presumed the probiotic intervention to some extent caused dysbiosis as inverse FMT. The role of probiotics in IBD requires further elucidation.

Advances in Preclinical In Vitro Models for the Translation of Precision Medicine for Cystic Fibrosis.

The development of preclinical in vitro models has provided significant progress to the studies of cystic fibrosis (CF), a frequently fatal monogenic disease caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR) protein. Numerous cell lines were generated over the last 30 years and they have been instrumental not only in enhancing the understanding of CF pathological mechanisms but also in developing therapies targeting the underlying defects in CFTR mutations with further validation in patient-derived samples. Furthermore, recent advances toward precision medicine in CF have been made possible by optimizing protocols and establishing novel assays using human bronchial, nasal and rectal tissues, and by progressing from two-dimensional monocultures to more complex three-dimensional culture platforms. These models also enable to potentially predict clinical efficacy and responsiveness to CFTR modulator therapies at an individual level. In parallel, advanced systems, such as induced pluripotent stem cells and organ-on-a-chip, continue to be developed in order to more closely recapitulate human physiology for disease modeling and drug testing. In this review, we have highlighted novel and optimized cell models that are being used in CF research to develop novel CFTR-directed therapies (or alternative therapeutic interventions) and to expand the usage of existing modulator drugs to common and rare CF-causing mutations.

T-cell activation and B-cell interaction signatures in rectal tissues are associated with HIV replication in ex-vivo model of infection.

The rectal mucosa is a critical site of HIV vulnerability. We sought to identify transcriptomic features of rectal mucosal tissue prior to exposure associated with support or restriction of HIV replication. Rectal tissue from HIV-negative cis gender men ( n = 57) underwent concurrent RNAseq transcriptomic analyses (two biopsies/participant) and challenge with HIV in the ex-vivo explant model of infection (three biopsies challenged/participant) as part of a larger cohort study to understand the rectal mucosal immune environment among MSM. P24 was quantified in the explant supernatants over a culture period of 18 days via ELISA. Participant median p24 log area under the curve was correlated with bulk transcriptomic data (Illumina HiSeq3000) to identify associations between gene expression and p24 production. Significant differentially expressed genes (DEGs) were identified via DESeq2 analysis and analyzed with Reactome to identify pathways of interest. In total, 183 DEG (181 upregulated, two downregulated) were associated with higher p24 accumulation in the ex-vivo challenge model, including T-cell activation, B-cell function, and chemokine DEG. Reactome analysis of the upregulated genes identified 'Adaptive Immune System', 'Cytokine Signaling in Immune System', and 'Innate Immune System' as significantly upregulated pathways. For the first time, we identified rectal tissue transcriptomic signatures associated with increased p24 production utilizing an ex-vivo model. Our findings are highly relevant to HIV transmission and the early establishment of HIV reservoirs in humans, and future studies should examine the identified pathways as targets for new or improved biomedical prevention or treatment interventions.

Characterization of the mechanism of Scutellaria baicalensis on reversing radio-resistance in colorectal cancer

Scutellaria baicalensis (SB) has been shown to improve the therapeutic effects of colorectal cancer (CRC) and perform well for reversing radio-resistance in different cancers. However, its potential function and mechanism related to radio-resistance in CRC has not been explored. A radio-resistant human CRC cell line (HCT116R) was applied. A network pharmacological analysis was performed to reveal the potential mechanism of SB for reversing radio-resistance in CRC, and computational pathological analysis was applied to indicate the clinicopathological significance of the key targets. Then, our hypothesis was further verified by molecular docking. The network pharmacology analysis showed that wogonin is the key compound of SB for reversing the radio-resistance of CRC. A Kyoto Encyclopedia of Genes and Genomes analysis showed that the genes for SB that reverse radio-resistance in CRC are mainly involved in steroid hormone biosynthesis. An enrichment analysis pointed out that Sulfotransferase family 2B member 1 (SULT2B1) is a potentially vital gene. SULT2B1 was demonstrated as being highly expressed in CRC and upregulated in radio-resistant rectal tissues or cell lines. A CCK-8 and clone formation test showed that the viability and clone formation ability of HCT116R were significantly decreased by wogonin combined with radiotherapy, compared to radiotherapy alone. By contrast, flow cytometry revealed that the apoptosis of HCT116R was significantly increased when wogonin treatment combined with radiotherapy, compared with radiotherapy alone. Molecular docking verification indicated that SULT2B1 and wogonin have a good binding ability. Taken together, SULT2B1 may be the potential drug target in treating radio-resistant CRC. Wogonin may be the core compound of SB for reversing radio-resistance in CRC by targeting SULT2B1.

Evaluation of histopathological response to neoadjuvant therapy in rectal cancer using slide-free, stain-free multimodal multiphoton microscopy.

Neoadjuvant therapy has become a standard treatment for patients with locally advanced rectal cancer to achieve better prognostic outcomes. The response to treatment has been shown to correlate closely with the prognosis. However, current evaluation systems only provide coarse assessment on limited information, due to the lack of accurate and reproducible approach for quantitation of different types of responses. In this study, a novel stain-free, slide-free multimodal multiphoton microscopy imaging technique was applied to image rectal cancer tissues after neoadjuvant therapies with high resolution and contrast. Qualitative and quantitative evaluation of tumor, stromal, and inflammatory responses were demonstrated which are consistent with current tumor regression grading system using American Joint Committee on Cancer criteria, showing the great potential of such approach to build a more informative grading system for accurate and standardizable assessment of neoadjuvant therapy in rectal cancer.

Role of islatravir in HIV treatment and prevention: an update.

To summarize recent updates on the potential role of islatravir for HIV treatment and prevention. Islatravir is an investigational antiretroviral agent with unique pharmacologic properties that facilitate flexible dosing regimens. Islatravir has demonstrated potent antiviral activity and a high barrier to resistance when combined with doravirine and lamivudine. A simplified two-drug HIV treatment regimen of islatravir combined with doravirine has also demonstrated comparable efficacy to standard of care three-drug regimens. The long half-life and high potency of islatravir's active metabolite may support its use as a long-acting option for HIV preexposure prophylaxis (PrEP). A once monthly oral dose of islatravir maintains effective concentrations of its active metabolite over the entire dosing interval. Furthermore, an investigational implantable formulation has been projected to provide efficacious concentrations for at least a year and exhibits comparable distribution into vaginal and rectal tissues making it a promising PrEP option for male and female individuals. Islatravir has minimal risks of drug interactions as it is not a substrate, inducer, or inhibitor of major drug metabolizers and transporters. Finally, clinical trials demonstrate islatravir's favorable safety profile revealing only mild and transient adverse events. Leveraging the unique pharmacological properties of islatravir offers opportunities for simplified HIV treatment regimens and long-acting PrEP making it a valuable addition to the antiretroviral arsenal.

Method and Critical Aspect of Semisolid Mixing

Semisolid preparations are widely used to deliver drugs through the skin, cornea, rectal tissue, nasal mucosa, vagina, buccal tissue, urethral membrane, and outer ear lining. They can prevent the first-pass metabolism, reduce side effects, provide immediate local effects, and increase patient compliance. However, an improper manufacturing process will produce a system with bad characteristics, one of which is the mixing process. Several conditions that need to be considered, such as vacuum, temperature, humidity, pressure, stirring speed, stirring time, shear stress, the volume of the mixture, and type of impeller, can affect the consistency, size, and dispersion of particle size, homogeneity, porosity, reactivity, and other characteristics that affect the quality of the semisolid system. Therefore, this article discusses the critical aspects of semisolid mixing, the types, principles, and specifications of several mixer tools and impellers, and how they affect the characteristics of semisolid systems. This review concludes that each type of semisolid preparation requires an impeller and mixer with the specifications and mixing conditions that suit the needs in maintaining the stability and quality of the semisolid system.

Role of Microbial Metabolites of Histidine in the Development of Colitis.

Colitis is a chronic relapsing inflammatory disease of colon. Clinical studies show that meat-rich diet plays a critical role in the relapse of colitis. However, it is unclear whether the microbial metabolites of histidine, which is an amino acid widely found in meat, have an impact on the health of the intestine. Six metabolites of histidine are given to IEC-6 cells. The cell activity measurement shows that imidazole propionate (IMP) is the most detrimental metabolite. Then, IMP is injected to mice by rectal administration, with blood and colon tissues collected for the measurement of colitis related parameters. The results show that treatment with IMP significantly increased NF-κB, iNOS, and IL-6, decreased number of goblet cell, and inhibited expressions of miR-146b. However, overexpression of miR-146b in mice rescues the decline of the physical condition. Additionally, Notch receptor 1 (Notch1) is identified as a target gene of miR-146b. Further analysis shows that miR-146b restored the abundance of goblet cells by regulating Notch1 signaling pathway. IMP is able to induce intestinal inflammation, impairs the intestinal barrier, and affects the proliferation of goblet cells. The underlined mechanism may partially contribute to the dysregulation of miR-146b/Notch1 axis.

The dynamic cellular and molecular features during the development of radiation proctitis revealed by transcriptomic profiling in mice

BackgroundRadiation proctitis (RP) is the most common complication of radiotherapy for pelvic tumor. Currently there is a lack of effective clinical treatment and its underlying mechanism is poorly understood. In this study, we aimed to dynamically reveal the mechanism of RP progression from the perspective of RNomics using a mouse model, so as to help develop reasonable therapeutic strategies for RP.ResultsMice were delivered a single dose of 25 Gy rectal irradiation, and the rectal tissues were removed at 4 h, 1 day, 3 days, 2 weeks and 8 weeks post-irradiation (PI) for both histopathological assessment and RNA-seq analysis. According to the histopathological characteristics, we divided the development process of our RP animal model into three stages: acute (4 h, 1 day and 3 days PI), subacute (2 weeks PI) and chronic (8 weeks PI), which could recapitulate the features of different stages of human RP. Bioinformatics analysis of the RNA-seq data showed that in the acute injury period after radiation, the altered genes were mainly enriched in DNA damage response, p53 signaling pathway and metabolic changes; while in the subacute and chronic stages of tissue reconstruction, genes involved in the biological processes of vessel development, extracellular matrix organization, inflammatory and immune responses were dysregulated. We further identified the hub genes in the most significant biological process at each time point using protein-protein interaction analysis and verified the differential expression of these genes by quantitative real-time-PCR analysis.ConclusionsOur study reveals the molecular events sequentially occurred during the course of RP development and might provide molecular basis for designing drugs targeting different stages of RP development.

Histopathological Comparison between Punch and Full Thickness Rectal Biopsy in Diagnosis of Hirschsprung’s Disease

Background: The diagnosis of Hirschsprung’s disease (HD) is dependent on the histological study of rectal ganglion cells. Open rectal biopsy is the mainstay that requires general anesthesia (GA) and carries risk of many complications. Rectal punch biopsy has gained wide acceptance and has become the choice as it is simple, safe, having no need of anesthesia, done bedside with a simple biopsy forcep with high degree of accuracy and virtual absence of any complications. Objective: To find out the efficacy of rectal punch biopsy in the diagnosis of Hirschsprung’s disease. Materials and Methods: A cross sectional descriptive observational study was carried out with the histological findings of 60 rectal punch biopsies with corresponding 60 full thickness rectal biopsies from 60 suspected HD patients from March 2018 to February 2020. Sections made from the paraffin blocks were stained with H&amp;E and were examined for the presence or absence of ganglion cells and hypertrophic nerve fibers in the submucosa. Then the relations were studied. These findings were compared with full thickness rectal biopsy. Result: There were 40(66.7%) cases of male and 20(33.3%) cases of female including 26 (43.3%) cases of infants. Out of 60 cases, 10 cases (seven inadequate and three suspicious) of punch biopsy were not compared with full thickness rectal biopsy. Out of 50 cases, 38(63.3%) cases were HD in punch biopsy; among these 36(92.3%) cases were HD and 2(18.2%) cases were Non-HD in full thickness rectal tissue biopsy. The accuracy of punch biopsy was measured by sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and efficiency of the test which were 92.3%, 81.8%, 94.7%, 75.0% and 90.0% respectively. Conclusion: The rectal punch biopsy is simple, safe, no need of anesthesia and can be done with a simple biopsy forcep with high degree of accuracy. KYAMC Journal Vol. 13, No. 01, April 2022: 37-40

Understanding the Interaction Between Prostaglandins and Toll-Like Receptors in Nicotine-Induced Rectal Tumor in Animals.

In the present study, we tried to understand the crosstalk between prostaglandins-COX-mediated rectal tumors and toll- like receptors in rats. The tumor was induced using nicotine (100 μL/mL). Following the induction, the serum and rectal tissue were analyzed for Lipo-polysaccharides (LPS) and prostaglandin E2 in serum, and tissue expression of inflammatory mediators like TLR2,4, NFkB; cancer markers like Matrix metalloproteases 2 (MMP2), 9 and Cyclo-oxygenases 2 (COX-2) were estimated. The gut microflora analysis was carried out using the fresh fecal samples of both the study groups. In nicotine-induced group, there was a significant alteration in the gut microflora toward high Gram-negative strains and a decline in Gram-positive populations. All the inflammatory as well as cancer prognostic markers were significantly increased in the tumor-induced animals. From the present study, it could be concluded that nicotine significantly induced rectal cancer in the mice model by modu- lating gut microflora and increasing COX-2 and prostaglandin E2 levels.

RIOK1 mediates p53 degradation and radioresistance in colorectal cancer through phosphorylation of G3BP2.

RIO Kinase 1 (RIOK1) is involved in various pathologies, including cancer. However, the role of RIOK1 in radioresistance of colorectal cancer (CRC) remains largely unknown. In this study, we reported that RIOK1 was overexpressed in rectal cancer tissue with weaker tumor regression after neoadjuvant chemoradiotherapy (neoCRT). Moreover, higher RIOK1 expression predicted a poor prognosis in patients with rectal cancer. Blockade of RIOK1 using Toyocamycin, a pharmacological inhibitor of RIOK1, or by knocking down its expression, decreased the resistance of CRC cells to radiotherapy in vitro and in vivo. A mechanistic study revealed that RIOK1 regulates radioresistance by suppressing the p53 signaling pathway. Furthermore, we found that RIOK1 and Ras-GAP SH3 domain binding protein 2 (G3BP2) interact with each other. RIOK1 phosphorylates G3BP2 at Thr226, which increases the activity of G3BP2. RIOK1-mediated phosphorylation of G3BP2 facilitated ubiquitination of p53 by murine double minute 2 protein (MDM2). Altogether, our study revealed the clinical significance of RIOK1 in CRC, and therapies targeting RIOK1 might alleviate the CRC tumor burden in patients.

Preoperative biomarkers related to inflammation may identify high-risk anastomoses in colorectal cancer surgery: explorative study.

BackgroundColorectal anastomotic leakage can be considered a process of failed wound healing, for which related biomarkers might be a promising research area to decrease leak rates.MethodsPatients who had elective surgery with a primary anastomosis for non-metastatic colorectal cancer, at two university hospitals between 1 January 2010 and 31 December 2015 were included. Patients with an anastomotic leak were identified and matched (1:1) to complication-free controls on the basis of sex, age, tumour stage, tumour location, and operating hospital. Preoperative blood samples were analysed by use of protein panels associated with systemic or enteric inflammation by proteomics, and enzyme-linked immunosorbent assays. Multivariable projection methods were used in the statistical analyses and adjusted for multiple comparisons to reduce false positivity. Rectal cancer tissue samples were evaluated with immunohistochemistry to determine local expression of biomarkers that differed significantly between cases and controls.ResultsOut of 726 patients undergoing resection, 41 patients with anastomotic leakage were matched to 41 controls. Patients with rectal cancer with leakage displayed significantly elevated serum levels of 15 proteins related to inflammation. After controlling for a false discovery rate, levels of C-X-C motif chemokine 6 (CXCL6) and C-C motif chemokine 11 (CCL11) remained significant. In patients with colonic cancer with leakage, levels of high-sensitivity C-reactive protein (hs-CRP) were increased before surgery. Local expression of CXCL6 and CCL11, and their receptors, were similar in rectal tissues between cases and controls.ConclusionPatients with anastomotic leakage could have an upregulated inflammatory response before surgery, as expressed by elevated serological levels of CXCL6 and CCL11 for rectal cancer and hs-CRP levels in patients with colonic cancer respectively.

Toll-like Interleukin -1 Receptor Regulator (TILRR) Protein, a Major Modulator of Inflammation, is Expressed in Normal Human and Macaque Tissues and PBMCs.

PurposeTILRR is a modulator of genes in the NF-κB inflammation pathway. It regulates inflammation-responsive genes, the secretion of inflammatory mediators, and the migration of immune cells. Because inflammation drives the pathogenesis of many infectious and inflammatory diseases, it is important to know the expression of TILRR protein in tissues and cells. This study examined TILRR protein expression in healthy adult human and macaques’ tissues and PBMCs (peripheral blood mononuclear cells).Methods and ResultsTissues (trachea, lungs, stomach, small intestine [ileum], cecum, colon, rectum, vagina, cervix, uterus, and penis) and PBMCs from humans and macaques were lysed in RIPA (radioimmunoprecipitation assay) lysis buffer. The TILRR protein was examined by fluorescent Western blot analysis. The relative fluorescence units (rfu) of TILRR protein expression were quantified by Image Studio software (LI-COR). The results showed that adult healthy female (n=1) rectal and cervicovaginal tissues expressed a higher level of TILRR protein than the other tissues (trachea, lungs, stomach, small intestine [ileum], cecum, colon, uterus, and penis) examined. Like humans, the lungs, colon, and rectal tissues of healthy adult female cynomolgus monkeys (Macaca fascicularis) (n=2) expressed the TILRR protein. In addition, PBMCs of healthy adult women (n=4), adult female cynomolgus monkeys (Macaca fascicularis) (n=4), and adult male and female rhesus monkeys (Macaca mulatta) (n=4) showed a similar expression level of TILRR protein (p= 0.2858). TILRR protein was not detected in most of the human cell lines examined, except in Jurkat cells.ConclusionOur study for the first time showed that TILRR protein is expressed in healthy adult human and monkey tissues and PBMCs. The TILRR protein in these tissues and PBMCs may play a role in the inflammatory response of these tissues and cells in response to infectious pathogens.

Expression profiles of three proteins in various cancerous tissues of digestive system

A variety of gastrointestinal tumors are the most malignancies with high morbidity and mortality globally. The previous studies show that there are abnormal expressions of TATA-box binding protein associated factor 1 like (<italic>TAF</italic>1<italic>L</italic>), stomatin-like protein 1 (<italic>STOML</italic>1) and stomatin-like protein 2 (<italic>STOML</italic>2) genes in oral, esophageal and colorectal cancer lesions. To explore the potential roles of above three genes in the carcinogenesis of digestive system, the differential expressions of TAF1L, STOML1 and STOML2 in cancerous and paracancerous tissues are compared and analyzed by multiple immunohistochemical staining using microarray of 6 cancer tissues of esophagus, stomach, liver, pancreas, colon and rectum with matched paracancerous tissues. The results show that STOML2 is significantly overexpressed in all of 6 gastrointestinal cancers <inline-formula><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M4"><mml:mo stretchy="false">(</mml:mo><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn mathvariant="normal">0.001</mml:mn><mml:mo stretchy="false">)</mml:mo></mml:math></inline-formula>, and STOML1, one of the same family member of STOML2, is highly expressed only in esophageal and liver cancers <inline-formula><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M5"><mml:mo stretchy="false">(</mml:mo><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn mathvariant="normal">0.005</mml:mn><mml:mo stretchy="false">)</mml:mo></mml:math></inline-formula>. In addition, except for pancreatic cancer, TAF1L is also abnormal highly expressed in other 5 cancer tissues <inline-formula><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M6"><mml:mo stretchy="false">(</mml:mo><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn mathvariant="normal">0.005</mml:mn><mml:mo stretchy="false">)</mml:mo></mml:math></inline-formula>. At the same lesion tissue section, STOML1, STOML2 and TAF1L proteins are colocated with the key protein MMP9 of epithelial-mesenchymal transformation in positive cells. This study indicates that the three proteins of TAF1L, STOML1 and STOML2 might have a wide range of cancer promoting effects in multiple cancer tissues in the digestive system.In addition, due to their different expression levels in a variety of cancer tissues, they have tissue specificity and different functional missions.Further exploration of the molecular pathological potential of TAF1L, STOML1 and STOML2 in the occurrence and development of malignant tumors in digestive system can make them serve as new molecular markers for early diagnosis, differential diagnosis, targeted therapy and prognosis prediction in multiple digestive tumors. <fig fig-type="abstract-image" id="F1" orientation="portrait" position="float"><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="1000-2618-2022-39-3-253/E85B5A24-8ED1-441e-8E1E-AB53DA3D3AE9-F001.jpg"/></fig>

Preparation, in vitro and in vivo Evaluation of Thermosensitive in situ Gel Loaded with Ibuprofen-Solid Lipid Nanoparticles for Rectal Delivery.

BackgroundIbuprofen (IBU), a nonsteroidal anti-inflammatory drug, shows poor gastrointestinal absorption due to its low solubility, which limits its clinical application.ObjectiveIn the present study, we aimed to develop thermosensitive gel-mediated ibuprofen-solid lipid nanoparticles (IBU-SLN-ISG) to improve the dissolution and bioavailability of IBU after rectal delivery.MethodsIBU-loaded SLNs (IBU-SLNs) were developed and optimized applying Box-Behnken design. The optimized IBU-SLNs were characterized by physicochemical parameters and morphology. Then, the optimized IBU-SLNs was incorporated into the gel and characterized for gel properties and rheology and investigated its release in vitro, pharmacokinetics in vivo, rectal irritation and rectal retention time.ResultsThe optimized SLNs had an EE of 90.74 ± 1.40%, DL of 11.36 ± 1.20%, MPS of 166.77 ± 2.26 nm, PDI of 0.27 ± 0.08, and ZP of −21.00 ± 0.59 mV. The FTIR spectra confirmed successful encapsulation of the drug inside the nanoparticle as only peaks responsible for the lipid could be identified. This corroborated well with XRD spectra, which showed a completely amorphous state of the IBU-SLNs as compared to the crystalline nature of the pure drug. The gelation temperature of the prepared IBU-SLN-ISG was 33.30 ± 0.78°C, the gelation time was 14.67 ± 2.52 s, the gel strength was 54.00 ± 1.41 s, and the mucoadhesion was (11.54±0.37) × 102dyne/cm2. The in vitro results of IBU-SLNs and IBU-SLN-ISG showed a biphasic release pattern with initial burst release followed by sustained release. More importantly, IBU-SLN-ISG produced much better absorption of IBU and improved bioavailability in rats. In addition, IBU-SLN-ISG caused no irritation or damage to rectal tissues, and could be retained in the rectum for a long time.ConclusionThermosensitive in situ gel loaded with IBU-solid lipid nanoparticles might be further developed as a more convenient and effective rectal dosage form.

Hypoxic Cell-Derived Extracellular Vesicles Aggravate Rectal Injury Following Radiotherapy via MiR-122-5p.

Radiation-induced rectal injury is a common side effect of radiotherapy. Hypoxia often occurs after radiotherapy. This study aimed to explore the bystander effect of hypoxia on radiation-induced rectal injury. In vivo, apoptosis increased nearby the highly hypoxic area in the rectal tissues in the mouse models of radiation-induced rectal injury, indicating the potential involvement of hypoxia. In vitro, flow cytometry and Western blotting showed that both hypoxia and hypoxic human intestinal epithelial crypt (HIEC) cell supernatant promoted apoptosis in normoxic HIEC cells. The pro-apoptotic effect of extracellular vesicles (EVs) derived from hypoxic HIEC cell to normoxic HIEC cells was then determined. MiR-122-5p was chosen for further studies through a microRNA (miRNA) microarray assay and apoptosis was alleviated in cells receiving miR-122-5p inhibiting hypoxic EVs. Together, our study demonstrated that the miR-122-5p containing-EVs derived from hypoxic HIEC cells promoted apoptosis in normoxic HIEC cells. Hypoxic EV-derived miR-122-5p plays a critical pathologic role in radiation-induced rectal injury and may be a potential therapeutic target.