Anal Tissue Research Articles

Topical Protease Inhibitor Increases Tumor-Free and Overall Survival in CD4-Depleted Mouse Model of Anal Cancer.

Patients with immunodeficiencies and older age are at an increased risk of anal cancer. Transgenic K14E6/E7 mice with established high-grade anal dysplasia were treated topically at the anus with the protease inhibitor saquinavir (SQV) in the setting of CD4+ T-cell depletion to mimic immunodeficiency. To ensure tumor development, specific groups were treated with a topical carcinogen (7,12-Dimethylbenz[a]anthracene (DMBA)). The treatment groups included the vehicle (control), DMBA only, topical SQV, and topical SQV with DMBA, as well as the same four groups with CD4 depletion. The mice were monitored weekly for tumor development. Upon reaching 20 weeks of treatment, the mice were sacrificed, and their anal tissue was harvested for histological analysis. None of the mice in the SQV or control groups developed overt anal tumors, except three mice that were CD4-depleted. The CD4-depleted mice treated with DMBA had significantly increased tumor-free survival and overall survival as well as decreased tumor-volume growth over time when treated with SQV. These data suggest that topical SQV, in the setting of CD4 depletion and high-grade anal dysplasia, can increase tumor-free and overall survival; thus, it may represent a viable topical therapy to decrease the risk of progression of anal dysplasia to anal cancer.

Isolation and genomic characterization of chicken infectious anemia virus in Jiangsu province of China during 2020-2022.

As an immunosuppressive disease virus, chicken infectious anemia virus (CIAV) mainly infects chickens, causing aplastic anemia and systemic lymphoid tissue atrophy. In recent years, the prevalence of CIAV in the poultry industry globally has caused huge economic losses. In this study, a total of 223 clinical samples, including anal swabs, tissues, blood, and vaccines, were collected from 19 broiler farms or breeding companies in Jiangsu province, with symptoms of significant anemia and immunosuppression during 2020-2022. Among them, 75 samples (75/223, 33.6%) were positive for CIAV in polymerase chain reaction (PCR) test, and 20 CIAV strains were successfully isolated. The phylogenetic trees of the 20 isolates and 42 CIAV strains deposited in GenBank formed four distinct groups (A-D). And the isolates mainly belonged to Group A but with high genetic diversity. Analysis for VP1 indicated that these isolates possess key characteristics of highly pathogenic strains. Meanwhile, VP2 and VP3 were much conserved with much fewer mutations compare to VP1. The above epidemiological study of CIAV provides novel insights into molecular characterization of CIAV and lays the foundation for developing efficient strategies for control of CIAV in China.

The beneficial effect of Sanhuang ointment and its active constituents on experimental hemorrhoids in rats

Ethnopharmacological relevanceSanhuang ointment (SHO) has been widely used in the traditional Chinese medical system for 1500 years and has efficacy in clearing away heat and dampness, reducing swelling, and alleviating pain. Hemorrhoids will damage the normal physiological function of the body, resulting in obstructed defecation, accompanied by massive hemorrhage and necrosis of tissues and cells, which is easy to breed bacteria and cause infection. SHO can promote lesion healing in hemorrhoid rats, but the pharmacological mechanism underlying this effect remains unknown. AimTo evaluate the effect of SHO on experimental hemorrhoids in rats induced by croton oil and glacial acetic acid. Materials and methodsIn this research, the effective components of SHO were analyzed in detail by High performance liquid chromatography (HPLC) and Liquid chromatography/mass spectrometry (LC/MS). Hemorrhoids were induced by 6% balsam and glacial acetic acid respectively in the anorectal region of rats. SHO was administered externally to the anorectal region of rats at doses of 185 mg/g (crude drug/ointment), 370 mg/g (crude drug/ointment) and 740 mg/g (crude drug/ointment) for 11 days. Mayinglong musk hemorrhoids ointment (1 g/kg) and Taining cream (1 g/kg) were used as reference anti hemorrhoids drugs. On the 11th day, hemorrhoids were evaluated by measuring the biochemical parameters of hemorrhoids in rats and the histology of anorectal tissues. ResultsUsing high performance liquid chromatography liquid chromatography mass spectrometry, 41 compounds, including phenylpropionic acids and alkaloids, were identified. the fingerprints of 18 common peaks were identified. In Hemorrhoids like rats, acetic acid induced inflammation was inhibited in a dose-dependent manner during SHO treatment. In addition, the detailed experimental results show that SHO can effectively improve hemorrhoids by inhibiting the production of inflammatory cytokines in serum, reversing the down-regulation of vanillin subtype 1 (TRPV1), calcitonin gene related peptide (CGRP) and substance P (SP) levels of pain related genes in anal tissues, and the up regulation of Vascular endothelial growth factor (VEGF) levels of vascular growth related genes. ConclusionThe results showed that SHO could alleviate the edema caused by the exudation of anorectal tissue fluid in rats by anti-inflammatory effect and reducing the Vascular permeability of rats. The study validates the traditional use of SHO in the treatment of hemorrhoids and demonstrates its anti-hemorrhoidal potential.

Hemorrhoid disease: A review on treatment, clinical research and patent data.

Hemorrhoid disease (HD) is an anal-rectal ailment that is commonly painful or may be painless and causes rectal bleeding with or without prolapsing anal tissue. It is generally associated with bleeding, prolapse, pruritus, and discomfort, which results in a diminished quality of life and well-being. To highlight the recent developments in terms of safety, clinical efficacy, and marketed formulation for the effective management of hemorrhoids. Reported literature available on Scopus, PubMed, Science Direct, Clinicaltrails.gov, and from many reputed foundations has been studied to summarize the recent development and clinical studies for the management of hemorrhoids. The high incidence of hemorrhoids obliges the development of new molecules; therefore, safe and efficient drugs to confer protection against hemorrhoids are urgently needed. This review article mainly focuses on the newer molecules to overcome hemorrhoids and also emphasizes various studies carried out in the past.

EFFECT OF JALAUKAVACHVARANA IN THROMBOSED HAEMORRHOID: A CASE STUDY

Haemorrhoids are a prevalent anorectal condition. It is of two types: Internal and external. A thrombosed haemorrhoid is a variety of external haemorrhoids in which there is painful swelling in anal tissue caused by a clot in a small vein of the anal skin. Case presentation: A 35 year old male patient visited OPD with complaints of protrusion of mass outside the anal region, constipation and pain during defecation. Routine clinical examinations and investigations are done. Diagnosis and management: Based on clinical examination, the case was diagnosed as thrombosed haemorrhoids and suggested for Jalaukavacharana on an OPD basis. The treatment aims to relieve the pain and reduce the inflammation. Outcome: There is marked improvement in thrombosed haemorrhoids. The inflammation is reduced with relief from pain. Discussion: Satisfactory results were obtained with a decrease in the size of haemorrhoids and relief from pain. This case highlights that Jalaukavacharana is a very effective treatment for thrombosed haemorrhoids.

Croton Oil-induced Hemorrhoidal Rats: Evaluation of Anti-inflammatory Mediated Anti-hemorrhoidal Activity of Tridax procumbens Whole Plant

Background: Hemorrhoids are anorectal disorders characterized by dilation of rectal blood vessels, inflammation, and prolapse of the anal tissue. The disease affects both men and women equally. In consequence, the increasing prevalence of the disease needs potential agents with efficiency and low cost to support the treatment. Objective: The treatment of hemorrhoids in modern medicine is still found to be in its infancy, as there is no precise treatment for the specific disease. Tridax procumbens Linn (T. procumbens) belongs to the family Asteraceae. From the literature, the plant was found to have a traditional use for the treatment of hemorrhoids. Hence, the present research was focused on evaluating the antiinflammatory mediated anti-hemorrhoidal potential of ethanolic whole plant extract of T. procumbens (EWETP) in croton oil-induced hemorrhoidal rats. Methods: An in vitro study was conducted to evaluate the percentage inhibition of protein (egg albumin) denaturation by Tridax procumbens. This study determined the initial protective role of plant extracts against inflammatory proteins. An in vivo, anti-inflammatory-mediated anti-hemorrhoidal study was carried out on T. procumbens in various groups of croton oil-induced hemorrhoidal rats. Wistar albino rats were selected for the present research, and hemorrhoids were induced in experimental animals using a croton oil preparation containing 6% croton oil, deionised water, pyridine, and diethyl ether. Pilex ointment was taken as a reference drug in this study. Inflammation seems to be a major pathway for the progression of hemorrhoids. Hence, in the present research, Evans blue (EB) extravasation technique was applied to quantify inflammatory proteins. On the last day of the study, blood samples were drawn from experimental animals to analyse serum and blood for TNF- α, IL-6, and the percentage of neutrophils count. Recto anal coefficient was calculated to measure croton oil-induced rectal inflammation in animals. Histopathological studies were carried out separately on a second set of animals to identify the protective role of T.procumbens on rectal tissue cell histology. Molecular docking studies were carried out to rule out the possible interaction of plant phytoconstituents with the COX-2 enzyme. Results: Results showed that intra rectal application of croton oil preparation in albino rats developed hemorrhoids by elevating serum TNF-α and IL-6 in positive control group animals compared to normal group rats. Treatment of albino rats with T. procumbens at doses of 200 mg/kg and 400 mg/kg in groups IV and V has shown a significant reduction in serum TNF-α and IL-6. Furthermore, in the study, T. procumbens exhibited a significant dose-dependent reduction of EB dye extravasation in study animals. The study observations also revealed the inhibitory effect of plant extract on the blood percentage of neutrophils count and recto anal coefficient compared to the positive control group rats. Histopathological studies conducted on rectoanal tissues showed the presence of minimal rectal tissue changes in plant extract-treated group animals compared to positive control group rats. Molecular docking studies explored the possible interaction of phenolic compounds of plant extract with the COX enzyme. Conclusion: It was concluded that Tridax procumbens had a protective role against inflammatory mediators in hemorrhoids. In hemorrhoidal rats, ethanolic leaf extract was found to reduce the plasma percentage of neutrophils and other inflammatory cytokines, TNF-α, and IL-6. It could be used as a therapeutic anti-inflammatory mediated anti-hemorrhoidal agent.

Vulval Fibroadenoma Presenting as a Unilateral Vulval Mass—A Report of a Common Neoplasm Present at an Uncommon Site

ABSTRACT Vulval fibroadenoma is a rare benign tumor arising from the ectopic breast tissue or mammary-like anal glandular tissue. Although it is a common breast neoplasm, its genital location makes it notable, with less than a hundred cases being reported in the literature so far. It is commonly known to present in the reproductive age group, similar to its breast counterpart, with its histogenesis being a frequent topic of discussion. Despite being benign and largely infrequent, these vulval neoplasms continue to be important reasons for medical consultation. We present one such case of vulval fibroadenoma presenting as an enlarged lymph node in a 37-year-old female.

Topical Protease Inhibitor Decreases Anal Carcinogenesis in a Transgenic Mouse Model of HPV Anal Disease.

Anal cancer is a major health problem. This study seeks to determine if the topical protease inhibitor Saquinavir (SQV), is effective at the prevention of anal cancer in transgenic mice with established anal dysplasia. K14E6/E7 mice were entered into the study when the majority spontaneously developed high-grade anal dysplasia. To ensure carcinoma development, a subset of the mice was treated with a topical carcinogen: 7,12-Dimethylbenz[a]anthracene (DMBA). Treatment groups included: no treatment, DMBA only, and topical SQV with/without DMBA. After 20 weeks of treatment, anal tissue was harvested and evaluated histologically. SQV was quantified in the blood and anal tissue, and tissue samples underwent analysis for E6, E7, p53, and pRb. There was minimal systemic absorption of SQV in the sera despite high tissue concentrations. There were no differences in tumor-free survival between SQV-treated and respective control groups but there was a lower grade of histological disease in the mice treated with SQV compared to those untreated. Changes in E6 and E7 levels with SQV treatment suggest that SQV may function independently of E6 and E7. Topical SQV decreased histological disease progression in HPV transgenic mice with or without DMBA treatment without local side effects or significant systemic absorption.

Conference report: Third annual young researcher forum of the Japan Society of Medical and Pharmaceutical Sciences for Traditional Medicine

AbstractFour young researchers presented their studies in the field of traditional Japanese herbal medicine (Wakan‐yaku) at the third annual young researcher forum entitled “Respecting and Breaking Traditions,” held 1 week before the 2022 annual meeting of the Japan Society of Medical and Pharmaceutical Sciences for Traditional Medicine. Dr. Yokogawa presented her research focusing on the mechanism of the Ayurveda drug‐eluting string, Kshara Sutra, in treating anal fistulas, reporting on its effects on the anal tissue in an anal fistula rat model and its antibacterial activity against Escherichia coli and Staphylococcus aureus, the causative organisms of anal fistulas. Mr. Nakamichi presented his research focusing on the relationship between the efficacy of Kampo medicines with anti‐obesity effects and intestinal microbiota composition. His results partially revealed that bofutsushosan and boiogito affect lipid metabolism in mice and correlate with the intestinal microbiota composition. Dr. Manse reported the anti‐inflammatory activity of diterpenoid components in Enmei‐so (Isodonis Herba), revealing its novel inhibitory activity on nitric oxide released from activated macrophages and on inflammatory cytokines, and identified active components contributing to the inhibition and the mechanism of action. Mr. Yoshikawa reported the anticancer activity of small molecular compounds isolated from the medicinal plants, Valeriana fauriei and Hypericum erectum. His results suggest that five compounds may increase the sensitivity of human cervical carcinoma cells to adriamycin via HSP105 expression suppression. The event was conducted online, and included 78 participants. We intend to hold and improve this forum in 2023 and the future.

Anal Cancer Prevention Through the Topical Use of Single or Dual PI3K/mTOR Inhibitors

IntroductionAnal dysplasia and anal cancer are major health problems. This study seeks to determine if inhibition of mTOR and/or PI3K pathways is effective at anal cancer prevention in mice with/without established precancerous lesions of the anus (anal dysplasia). MethodsK14E6/E7 mice were entered into the study at 5 wk, 15 wk, or 25 wk of age. Mice were treated with a topical carcinogen, 7,12-Dimethylbenz[a]anthracene (DMBA), which ensures carcinoma development within 20 wk. Treatment groups included: no treatment, DMBA only, topical Pictilisib (PI3K inhibitor) with/without DMBA, topical Sapanisertib (mTOR inhibitor) with/without DMBA, and topical Samotolisib (dual PI3K/mTOR inhibitor) with/without DMBA. Mice underwent weekly observations for anal tumor development (tumor-free survival). After 20 wk of treatment, anal tissue was harvested and evaluated histologically for squamous cell carcinoma (SqCC). ResultsAll topical treatments in conjunction with DMBA increased tumor-free survival in mice that started treatment at 15 wk of age when compared to DMBA-only treatment, except for Pictilisib + DMBA in males. Topical Sapanisertib increased tumor-free survival in mice regardless of starting treatment age. When examining tissue for microscopic evidence of SqCC, only topical Samotolisib in males decreased SqCC in the 15 wk starting mice. ConclusionsSapanisertib, the mTOR inhibitor, had the greatest effect, in terms of increasing tumor-free survival, regardless of starting time point or sex. Unlike the other treatments, Samotolisib, the dual PI3K/mTOR inhibitor, decreased microscopic evidence of SqCC when starting treatment at 15 wk of age but only in male mice.

The use of a topical protease inhibitor, Saquinavir, to alleviate mouse papillomavirus-mediated anal disease

Select protease inhibitors (PI) have been found to be effective in decreasing human papillomavirus oncoprotein expression. This study evaluated whether the topical PI, Saquinavir (SQV), promotes viral clearance in an infectious mouse model with Mus musculus papillomavirus 1 (MmuPV1). NOD scid gamma (NSG) mice were anally infected with ∼4 × 108 viral genome equivalents of MmuPV1 and 120 days post-infection (when majority have high-grade anal dysplasia), began topical treatments: control (mock), 7,12-dimethylbenz(a)anthracene (DMBA) only, once weekly to promote carcinogenesis, 1% SQV only, daily (Monday - Friday), and SQV + DMBA. Viral MmuPV1 load was analyzed from anal lavages pre and post-treatment. Anal tissue was harvested, processed, and evaluated for drug absorption, grade of anal disease, and anal viral RNA. Results suggest that topical SQV promotes decreased viral shedding in female mice treated with SQV.

Fulvestrant, an estrogen receptor inhibitor, relieves postoperative hemorrhoid edema via up-regulation of miR- 424-5p

Purpose: To investigate estrogen receptor (ER) expression in postoperative hemorrhoid edema tissues, and the likely mechanism involved in fulvestrant-mediated reduction of postoperative hemorrhoid edema.Methods: One hundred and eighty-five patients admitted to Jinshan Hospital of Fudan University, Shanghai who accepted hemorrhoidectomy were enrolled in this study. Primary cells were extracted from the anal margin tissues of patients for the determination of changes in ERα and vascular endothelial growth factor (VEGF). In vitro cellular experiments were performed in primary vascular endothelial cells to verify whether ER promoted postoperative perianal edema via the miR-424-5p estrogen receptor α gene (ESR1) axis. The cells were exposed to Fulvestrant, estradiol, and miR-424-5p mimic. Changes in expressions of ERα and VEGF were determined.Results: Fourteen patients (7.57 %), comprising 2 males (2.60 %) and 12 females (11.1 %), developedpostoperative anal margin edema. There was a significant difference in the incidence of postoperative anal edema between males and females (p < 0.05). Both immunohistochemistry and immunoblotting revealed markedly higher ERα levels in postoperative anal edema tissues than in preoperative tissues (p < 0.05). Moreover, ERα level was regulated by estradiol, and miR-424-5p targeted the estrogen receptor α gene (ESR1).Conclusion: Estradiol inhibits miR-424-5p through ERα in perianal tissues after hemorrhoid surgery. It increases VEGF and promotes perianal edema. However, fulvestrant inhibits ERα, thereby reducing VEGF expression and mitigating postoperative hemorrhoid edema, and therefore, has potential application for the management of postoperative hemorrhoid edema.

Investigation of different wavelengths for scattering-based light sheet microscopy.

Scattering-based light sheet microscopy (sLSM) is a microscopy technique that can visualize cellular morphologic details based on the scattering signal. While sLSM was previously shown to image animal tissues ex vivo at a cellular resolution, the wavelength used was chosen based on other in vivo microscopy technologies rather than through a comparison of the sLSM imaging performance between different wavelengths. In this paper, we report the development of a multi-wavelength sLSM setup that facilitates the investigation of different wavelengths for sLSM imaging. Preliminary results of imaging human anal tissues ex vivo showed that the sLSM setup allowed for comparisons of the cellular imaging performance at the same tissue location between different wavelengths. Both the quantitative analysis of the image contrast and the visual assessment by a pathologist showed that the imaging depth increased with wavelength, and the imaging depth increase was most notable around 600 nm. The preliminary results showed that the multi-wavelength sLSM setup could be useful in identifying the optimal wavelength for the specific tissue type.

Depo Medroxyprogesterone (DMPA) Promotes Papillomavirus Infections but Does Not Accelerate Disease Progression in the Anogenital Tract of a Mouse Model.

Contraceptives such as Depo-medroxyprogesterone (DMPA) are used by an estimated 34 million women worldwide. DMPA has been associated with increased risk of several viral infections including Herpes simplex virus-2 (HSV-2) and Human immunodeficiency virus (HIV). In the current study, we used the mouse papillomavirus (MmuPV1) anogenital infection model to test two hypotheses: (1) contraceptives such as DMPA increase the susceptibility of the anogenital tract to viral infection and (2) long-term contraceptive administration induces more advanced disease at the anogenital tract. DMPA treatments of both athymic nude mice and heterozygous NU/J (Foxn1nu/+) but ovariectomized mice led to a significantly increased viral load at the anogenital tract, suggesting that endogenous sex hormones were involved in increased viral susceptibility by DMPA treatment. Consistent with previous reports, DMPA treatment suppressed host anti-viral activities at the lower genital tract. To test the impact of long-term contraceptive treatment on the MmuPV1-infected lower genital tract, we included two other treatments in addition to DMPA: 17β-estradiol and a non-hormone based contraceptive Cilostazol (CLZ, Pletal). Viral infections were monitored monthly up to nine months post infection by qPCR. The infected vaginal and anal tissues were harvested and further examined by histological, virological, and immunological analyses. Surprisingly, we did not detect a significantly higher grade of histology in animals in the long-term DMPA and 17β-estradiol treated groups when compared to the control groups in the athymic mice we tested. Therefore, although DMPA promotes initial papillomavirus infections in the lower genital tract, the chronic administration of DMPA does not promote cancer development in the infected tissues in our mouse model.

Modern technologies in surgical treatment of combined anorectal pathology and their results

Surgery of combined anal and rectal diseases requires an unconventional tactics and methods of treatment of these pathologies, which provide minimal impact on tissues, preventing postoperative complications. The aim of the study was to conduct a comparative evaluation of the effectiveness of new radiosurgical and electrosurgical technologies in the treatment of combined anal and rectal diseases by analyzing the clinical course of the postoperative period and the impact of these energy devices on the morphostructure of operated tissues. The results of treatment of 747 patients with combined anal and rectal diseases divided into four research groups and control group have been analyzed. In the first research group 169 (22,6%) patients with electrosurgery device “ERBE ICC 200” have been operated on in the second research group – 114 (15,3%) patients with electrosurgery device "EFA", in the third research group – 107 (14,3%) patients – with electrosurgery device "KLS Martin" and in the fourth research group – 245 (32,8%) patients with radiosurgery device "Surgitron". The control group consisted of 112 (14,9%) patients operated with a surgical scalpel. After surgery 30 patients from each study group underwent morphological investigation of rectal and anal tissues for measurement of the thickness of thermal impacts on them, which was performed using an eyepiece-micrometer scale. Due to the minimal and insignificant impact on the tissues during application of electrosurgery devices “KLS Martin”, “EFA” and “ERBE ICC 200”, as well as radiosurgery device "Surgitron", there were detected neither scar anal strictures in any of the four study groups, nor scarring deformities of the pararectal areas, which contributed to the cosmetic nature of the combined operations and caused rapid rehabilitation of patients in the study groups. In the control group in 2 (2%) patients the formation of scar anal stricture was diagnosed, which required conservative (1 patient) and operative (1 patient) measures to eliminate them. Using modern radiosurgery and electrosurgery technologies for treatment of combined anal and rectal diseases reduces the operation duration, volume of bleeding and intensity of the postoperative pain. Application of these above technologies prevents the occurance of anal strictures and scar pararectal deformations.

Immunogenicity of an inactivated novel goose parvovirus vaccine for short beak and dwarfism syndrome in Cherry Valley ducks.

Duck short beak and dwarfism syndrome (SBDS) is a viral infectious disease caused by novel goose parvovirus (NGPV), which has been responsible for serious economic losses to the Chinese duck industry in recent years. Currently, there is no effective vaccine against this disease. In this study, we developed an inactivated virus vaccine candidate for SBDS based on NGPV strain DS15 isolated from a duck in China. Immune efficacy was evaluated in 112 ducks, which were randomly divided into vaccination, challenge-control, vaccination-challenge, and blank control groups (28 per group). Clinical characteristics, antibodies, virus excretion, viremia, and pathological changes were monitored. No morbidity or death was observed in the immunized ducks, which showed normal weight and a good mental state. High levels of serum antibodies (optical density at 450nm of ~0.63) were detected in ducks immunized with the inactivated vaccine at 7days post-vaccination (dpv), and the titer of virus-neutralizing antibodies increased from 1:23 to 1:28.5 from 7 to 42dpv. Measurement of the viral load in anal swab, serum, and tissue samples showed that vaccination significantly inhibited the replication of NGPV in immunized ducks. Moreover, NGPV could not be isolated from the spleens of immunized or vaccinated and challenged ducks. Collectively, these results demonstrate that the newly developed inactivated NGPV vaccine, administered in an oil emulsion adjuvant, possesses good immunogenicity and represents a potentially powerful tool for SBDS prevention and control.

Systemic Delivery of a Dual PI3K/mTOR Inhibitor More Effective than Topical Delivery in Preventing Anal Carcinogenesis in an HPV Transgenic Mouse Model.

Anal dysplasia is a growing health concern that over time can result in squamous cell carcinoma (SqCC) of the anus. In this study, we compare a topical versus systemic (oral) administration of LY3023414, a dual PI3K/mTOR inhibitor, to prevent anal carcinogenesis in a Human Papillomavirus (HPV) mouse model of anal cancer. K14E6/E7 transgenic mice were used to model HPV-induced anal carcinogenesis. Mice with varying starting anal histologies (normal histology, low-grade, and high-grade anal dysplasia) were treated topically at the anus or systemically via oral gavage with LY3023414 with or without topical carcinogen for 20 weeks. Mice were monitored for overt anal tumor development and anal tissue was assessed for histology and markers of PI3K and mTOR activity (pAKT and pS6, respectively). LY3023414 treatment, regardless of the mode of delivery, significantly decreased overt tumor development in mice starting with normal histology and low-grade anal dysplasia. Systemic LY3023414 treatment was more effective in delaying tumor onset than topical treatment. Mice treated with systemic LY3023414 had significantly reduced rates of anal SqCC when starting with normal and low-grade anal dysplasia compared to topical treatment. Topical treatment was only effective in reducing SqCC in the setting of low-grade dysplasia. LY3023414 inhibition of pAKT and pS6 expression varied with starting histology. Neither treatment mode was effective in the setting of high-grade anal dysplasia. Systemic LY3023414 treatment was more effective than topical application in delaying the progression of normal anal histology and low-grade dysplasia to anal cancer in HPV-associated mice.

Genome-wide host methylation profiling of anal and cervical carcinoma.

HPV infection results in changes in host gene methylation which, in turn, are thought to contribute to the neoplastic progression of HPV-associated cancers. The objective of this study was to identify joint and disease-specific genome-wide methylation changes in anal and cervical cancer as well as changes in high-grade pre-neoplastic lesions. Formalin-fixed paraffin-embedded (FFPE) anal tissues (n = 143; 99% HPV+) and fresh frozen cervical tissues (n = 28; 100% HPV+) underwent microdissection, DNA extraction, HPV genotyping, bisulfite modification, DNA restoration (FFPE) and analysis by the Illumina HumanMethylation450 Array. Differentially methylated regions (DMR; t test q<0.01, 3 consecutive significant CpG probes and mean Δβ methylation value>0.3) were compared between normal and cancer specimens in partial least squares (PLS) models and then used to classify anal or cervical intraepithelial neoplasia-3 (AIN3/CIN3). In AC, an 84-gene PLS signature (355 significant probes) differentiated normal anal mucosa (NM; n = 9) from AC (n = 121) while a 36-gene PLS signature (173 significant probes) differentiated normal cervical epithelium (n = 10) from CC (n = 9). The CC progression signature was validated using three independent publicly available datasets (n = 424 cases). The AC and CC progression PLS signatures were interchangeable in segregating normal, AIN3/CIN3 and AC and CC and were found to include 17 common overlapping hypermethylated genes. Moreover, these signatures segregated AIN3/CIN3 lesions similarly into cancer-like and normal-like categories. Distinct methylation changes occur across the genome during the progression of AC and CC with overall similar profiles and add to the evidence suggesting that HPV-driven oncogenesis may result in similar non-random methylomic events. Our findings may lead to identification of potential epigenetic drivers of HPV-associated cancers and also, of potential markers to identify higher risk pre-cancerous lesions.

Comparative characteristics of the results of surgical treatment of combined anorectal pathology using modern radiosurgical and high-frequency electrosurgical technologies

Annotation. Surgery of combined anal canal and rectal diseases requires an unconventional tactics and methods of treatment of these pathologies, which provide minimal impact on tissues, preventing postoperative complications. The purpose of the study was conduction a comparative evaluation of the results of surgical treatment of combined anorectal pathology using modern radiosurgery and electrosurgery technologies by analyzing the clinical course of the postoperative period and studying the depth of impact of these technological devices on the structure of operated tissues. The results of treatment of 635 patients with combined diseases of the anal canal and rectum, divided into four study groups, were analyzed. The control group consisted of 112 patients operated with a surgical scalpel. After surgery 30 patients from each study group underwent morphological investigation of rectal and anal tissues for measurement of the thickness of thermal impacts on them, which was performed using an eyepiece-micrometer scale. Statistical analysis of the obtained data was performed using IBM SPSS STATISTICS SUBSCRIPTIONAL TRIAL software. License number: L-CZAA-BHG85V. The following are used in the work: Kolmogorov-Smirnov test with Lilliefors and Shapiro-Wilk correction, Levene's criterion at p&lt;0.05; applied one-way analysis of variance, Bonferroni test; used the Welch test and the Brown-Forsyth test, the Games-Howell test; used the Kruskal-Wallis test, the Mann-Whitney test. It is established that due to the minimal and shallow impact on the tissues during application of electrosurgery devices “KLS Martin”, “EFA” and “ERBE ICC 200”, as well as radiosurgery device “Surgitron”, there were no detected neither scar anal strictures in any of the four study groups nor scarring deformities of the pararectal areas, which contributed to the cosmetic nature of the combined operations and caused rapid rehabilitation of patients in the study groups. In the control group in 2 (1,8%) patients diagnosed the formation of scar anal stricture which required conservative (1 patient) and operative (1 patient) measures to eliminate them. Using of modern radiosurgical and electrosurgical technologies for the treatment of combined anal and rectal diseases reduces the duration of surgery, intraoperative blood loss, postoperative pain and prevents scar strictures of the anal canal.

Sex-dependent development of Kras-induced anal squamous cell carcinoma in mice.

Anal squamous cell carcinoma (SCC) will be diagnosed in an estimated 9,080 adults in the United States this year, and rates have been rising over the last several decades. Most people that develop anal SCC have associated human papillomavirus (HPV) infection (~85–95%), with approximately 5–15% of anal SCC cases occurring in HPV-negative patients from unknown etiology. This study identified and characterized the Kras-driven, female sex hormone-dependent development of anal squamous cell carcinoma (SCC) in the LSL-KrasG12D; Pdx1-Cre (KC) mouse model that is not dependent on papillomavirus infection. One hundred percent of female KC mice develop anal SCC, while no male KC mice develop tumors. Both male and female KC anal tissue express Pdx1 and Cre-recombinase mRNA, and the activated mutant KrasG12D gene. Although the driver gene mutation KrasG12D is present in anus of both sexes, only female KC mice develop Kras-mutant induced anal SCC. To understand the sex-dependent differences, KC male mice were castrated and KC female mice were ovariectomized. Castrated KC males displayed an unchanged phenotype with no anal tumor formation. In contrast, ovariectomized KC females demonstrated a marked reduction in anal SCC development, with only 15% developing anal SCC. Finally, exogenous administration of estrogen rescued the tumor development in ovariectomized KC female mice and induced tumor development in castrated KC males. These results confirm that the anal SCC is estrogen mediated. The delineation of the role of female sex hormones in mediating mutant Kras to drive anal SCC pathogenesis highlights a subtype of anal SCC that is independent of papillomavirus infection. These findings may have clinical applicability for the papillomavirus-negative subset of anal SCC patients that typically respond poorly to standard of care chemoradiation.