Transcriptome analysis of eutopic endometrium in adenomyosis after GnRH agonist treatment

Background Adenomyosis is a chronic gynecological disease characterized by invasion of the uterine endometrium into the muscle layer. In assisted reproductive technology (ART), gonadotropin-releasing hormone agonist (GnRHa) is often used to improve pregnancy rates in patients with adenomyosis, but the underlying mechanisms are poorly understood. Methods Eutopic endometrial specimens were collected from patients with adenomyosis before and after GnRHa treatment in the midsecretory phase. RNA sequencing (RNA-Seq) of these specimens was performed for transcriptome analysis. The differentially expressed genes (DEGs) of interest were confirmed by real-time PCR and immunohistochemistry. Results A total of 132 DEGs were identified in the endometrium of patients with adenomyosis after GnRHa treatment compared with the control group. Bioinformatics analysis predicted that immune system-associated signal transduction changed significantly after GnRHa treatment. Chemokine (C-C motif) ligand 21 (CCL21) was found to be highly expressed in the eutopic endometrium after GnRHa treatment, which may be involved in the improvement of endometrial receptivity in adenomyosis. Conclusion This study suggests that molecular regulation related to immune system-associated signal transduction is an important mechanism of GnRHa treatment in adenomyosis. Immunoreactive CCL21 is thought to regulate inflammatory events and participate in endometrial receptivity in adenomyosis.

Retrospective analysis of OCT on MB characteristics and 1-year follow-up of the ISR incidence after the DES implantation in patients with MB

We used optical coherence tomography (OCT) to analyze the "half-moon" like phenomenon and its characteristics and observe 1-year follow-up of the in-stent restenosis (ISR) incidence after the drug eluted stent (DES) implantation in patients with the myocardial bridge (MB). Patients were retrospectively analyzed from January 2013 to December 2019. We used OCT to check 45 patients with MB and found a visible muscle layer (VML) around the vessel adventitia with the same or high density compared to the vessel media layer. There was not any significant difference in maximal thickness, maximal arch, and total length between the half-moon layer and the visible muscle layer groups (p > 0.05). Maximal thickness, arch, and total length of the half-moon layer were significantly positively related to VML, respectively (r = 0.962, 0.985, 0.742, p < 0.01). Of these 626 patients with MB seen by OCT, only 300 could be checked out by coronary angiography (CAG). Besides, the larger the thickness and arch of the VML around the vessel adventitia, the more severe the MB in these patients (p < 0.05). After the OCT use, there was no coronary perforation in these patients with MB covered with DES. After 1-year follow-up, ISR in MB covered with DES showed a notable difference among no MB, mild MB, moderate MB, and severe MB groups (p < 0.05), and ISR in DES aggravated with the MB severity. However, ISR in MB with and without covered with DES had no significant difference among the 4 groups (p > 0.05). OCT could evaluate MB characteristics accurately compared to IVUS and had a higher rate of detecting MB than CAG. Moreover, it is safe and effective to guide DES covering the mild MB segment in patients with severe coronary lesions detected by the OCT.

Pathomorphological changes in the early postnatal period in a calf with abomasum rupture

The main cause of the disease and death of calves in the early postembryonic period of development is a feeding disorder. The aim of the work was to analyze macro-and micromorphological changes in the digestive system of a calf with a rennet rupture. To achieve this goal, standard macro-and microscopic research methods were used. Macroscopic examination revealed thickening, erosion of the mucous membrane of the esophageal gutter rollers, accumulation of contents in the scar, mesh and book, hyperemia of the mucous membranes; thinning and rupture of the rennet wall; pronounced hyperemia of the mucous membrane of the small intestine, thickening of the mucous membrane of the thick section; an increase in mesenteric lymph nodes and a change in their consistency. Microscopically, the thickness of the epithelium, submucosal and muscle layer is most developed in the abomasum and was, respectively, 0.68-0.72 mm, 0.23-0.32 mm and 0.98-1.05 mm. Villi were found in the folds of the scar, numerous well – developed longitudinally oriented protrusions were found in the book, lymphocellular clusters and bottom glands were found in the rennet; folds with well – developed crypts were found in the colon. Thus, as a result of feeding coarse feed, the deceased calf has catarrhal erosive lesions of the esophageal trough; serous-catarrhal abomasitis with signs of hemorrhagic with dilation and perforation in the cardiac part; hemorrhagic omasitis against the background of a book blockage; catarrhal reticulitis; catarrhal ruminitis and catarrhal enterocolitis.

Spatiotemporal Expression of SHH/GLI Signaling in Human Fetal Bladder Development

Objectives: Sonic hedgehog (SHH) signaling is important in bladder development. Mice with defective hedgehog signaling develop bladder anomalies. Clinically, urinary tract malformations are reported in human fetuses and infants with mutations of SHH and related signaling pathway genes. Information on the expression of SHH and associated signaling genes in normal human bladder development is fragmentary. This study determined the temporal and spatial expression patterns of SHH signaling pathway components in human fetal bladders by immunohistochemistry (IHC).Material and Methods: Twenty-four bladder specimens from 16 male and 8 female human fetuses aged 12- to 36-week (wk) were obtained from the First Affiliated Hospital of Xi'an Jiaotong University. The tissue slides were processed for IHC staining with SHH, Patched1 (PTC-1), Patched2 (PTC-2), Smoothened (SMO), GLI1 and proliferating cell nuclear antigen (PCNA). The expression levels of each gene were analyzed by semi-quantitative histological scoring system.Results: High intensity of SHH and SMO expression was detected in developing bladder urothelial cells, with no staining in lamina propria (LP), but with minimal expression of SMO in differentiating smooth muscle (SM) layers. The spatial distribution pattern of PTC1 and GLI1 was more complex with minimal expression in the LP layer, moderate expression in the SM layer, and high expression in the urothelium. PTC2 expression was mainly localized in the urothelium and LP, but no expression in the SM layer. All of the SHH signaling components were detected in fetal bladder tissues throughout the development, with expression peaks at 12- and 23-wk, coinciding with high cell proliferation as indicated by PCNA staining in the cell nuclei of urothelium and SM.Conclusions: The autocrine SHH signaling in the developing urothelium, and paracrine SHH signaling in the developing smooth muscle layer, mediated by SMO, PTC-1 and GLI1 were demonstrated during human bladder development. Expression of SHH signaling components peaked at 12-and 23-wk. The first expression peak at 12-wk may relate to urothelium growth, SM induction, and dilation of the bladder cavity. The second expression peaked at 23-wk may relate to urothelium and SM layer differentiation.

Radical Radiation Therapy for Radiation-Induced Angiosarcoma with Local Control

Angiosarcoma is believed to be refractory to radical radiotherapy. Although no effective treatment for radiation-induced angiosarcoma has been established, surgery is generally chosen. We report a case of unresectable radiation-induced angiosarcoma after breast cancer surgery that responded remarkably to radical radiotherapy alone. A 78-year-old Japanese female had a large mass in her right breast that was diagnosed as angiosarcoma after biopsy and considered to be a radiation-induced angiosarcoma because the patient had been treated with postoperative irradiation to the right breast cancer 8 years earlier. There were no findings suggestive of lymphadenopathy or distant metastasis. Since the tumor was widely infiltrated to the skin and the muscle layer, radical radiotherapy (total dose 66 Gy) was performed. At this radiotherapy completion, the tumor was remarkably reduced, and there were no acute adverse effects except for grade 2 dermatitis. Pazopanib was started after radiotherapy but was discontinued since necrosis of the wound with hemorrhage was observed. External pneumothorax occurred due to the marked tumor shrinkage, but it was cured by conservative care. Although angiosarcoma is well known to be radioresistant, excellent local control may be obtained by radical radiotherapy in some cases. Radiotherapy should be considered as a treatment option in inoperable cases.

nPTD classification: an updated classification of gastric cancer location for function preserving gastrectomy based on physiological lymphatic flow

Background The correlation between tumor location and lymphatic flow distribution in gastric cancer has been previously reported, and PTD (Proximal – Transitional – Distal) classification was proposed. Our group updated and developed the nPTD classification. Method We retrospectively studied gastric cancer patients who underwent the dye method sentinel node biopsy from 1993 to 2020. The inclusion criteria were a single lesion type 0 cancer of ≤5 cm in the long axis, clinically node-negative, and invasion within the proper muscle layer pathologically. In this study, the distribution of dyed lymphatic flow was evaluated for each occupied area of the tumor. Results We included 416 patients in this study. The tumors located in the watershed of the right and left gastroepiploic arteries near greater curvature had extensive lymphatic flow; therefore, a newly circular region with a diameter of 5 cm is set on the watershed of the greater curvature between P and T zone as the ‘n’ zone. In addition, for cancers located in the lesser P curvature, lymphatic flow to the greater curvature was not observed. Therefore, the P zone was divided into two: the lesser curvature side (PL) and the greater curvature side (PG). Conclusions The advantage of the nPTD classification is that it provides not only proper nodal dissection but also adequate function-preserving gastrectomy. If the tumor is localized within the PL, the proximal gastrectomy resection area can be further reduced. In contrast, for cancers located in the ‘n’ zone, near-total gastrectomy is required because of the extensive lymphatic flow.

Prebiotic and synbiotic effect on rumen papilla length development and rumen pH in 12-week-old calves

Background and Aim: Europe and the USA have banned antibiotics use as growth promoters. There is a need for alternative products that can ensure production and health protection. Prebiotics has been proposed as alternatives because these materials have wide-ranging physiological effects on gut function, activity of the large intestinal microflora, mineral absorption, and immunity. The aim of this study was to determine the effect of three different doses of inulin, a prebiotic, in combination with probiotic Enterococcus faecium (a new synbiotic) on postnatal rumen development by comparing rumen papilla length, width, muscle layer thickness, and content pH level. Materials and Methods: Randomly selected 23 (±5)-days-old healthy male Holstein crossbreed calves, weighing 50 kg (±5 kg), were randomly allocated to seven groups, ten in each group. The calves were kept in a pen of 5, under the same conditions and were fed twice a day, ∼3.5 liters of whole milk per feeding. Control group (C n=10) was fed with whole milk only (no additives were added). The six other groups (three prebiotics and three synbiotics) received food additives with their morning milk feeding. The source of prebiotics, Jerusalem artichoke powder concentrate (JAPC) contained 50% of inulin. JAPC in doses of 6 g, 12 g, or 24 g were added to the milk. Formed prebiotic groups were denoted as PreG6, PreG12, and PreG24. To evaluate if the addition of the probiotic E. faecium 2×109 colony forming unit g–1 to manufacturer recommended dose of 0.25 g improves inulin effect on rumen, it was added to all their JAPC doses. The new content synbiotic groups were denoted as SynG6, SynG12, and SynG24. On day 57 of the study, when all calves were approximately 12 weeks old, they were slaughtered in a certified slaughterhouse. Tissue cultures for histological analysis were obtained from Saccus dorsalis and Saccus ventralis of the rumen. Tissue culture staining for histology was carried out using hematoxylin and eosin staining method. Rumen histological samples were used to measure papilla length, width, and muscle layer thickness. Each sample was used to make five measurements on the present rumen papilla. Results: The results showed that by adding 12 g of inulin to whole milk when feeding calves improves rumen papilla development, which is seen by increased length and width of papilla, especially in the Saccus ventralis region. By combing this dose of inulin with 0.25 g of E. faecium, a significant increase of papilla is achieved. Saccus ventralis muscle layer in the rumen is thicker than it is in Saccus dorsalis regardless of addition of prebiotics or synbiotics. Conclusion: The addition of inulin to whole milk can influence the pH of the rumen by making it more alkaline. The addition of prebiotic inulin and a novel synbiotic (inulin combined with E. faecium) can accelerate postnatal rumen development and improve its functionality.

Visceral organ morphogenesis via calcium-patterned muscle contractions

How organs achieve their final shape is a problem at the interface between physics and developmental biology. Organs often involve multiple interacting tissue layers that must be coordinated to orchestrate the complex shape changes of development. Intense study has uncovered genetic and physical ingredients driving the form of monolayer tissue. Yet, tracing dynamics across tissue layers and across scales -- from cell to tissue, to entire organs -- remains an outstanding challenge. Here, we study the midgut of Drosophila embryos as a model visceral organ to reconstruct in toto the dynamics of multi-layer organ formation in vivo. Using light-sheet microscopy, genetics, computer vision, and tissue cartography, we extract individual tissue layers to map the time course of shape across scales. We identify the kinematic mechanism driving the shape change due to tissue layer interactions by linking out-of-plane motion to active contraction patterns, revealing a convergent extension process in which cells deform as they flow into deepening folds. Acute perturbations of contractility in the muscle layer using non-neuronal optogenetics reveals that these contraction patterns are due to muscle activity, which induces cell shape changes in the adjacent endoderm layer. This induction cascade relies on high frequency calcium pulses in the muscle layer, under the control of hox genes. Inhibition of targets of calcium involved in myosin phosphorylation abolishes constrictions. Our study of multi-layer organogenesis reveals how genetic patterning in one layer triggers a dynamic molecular mechanism to control a physical process in the adjacent layer, orchestrating whole-organ shape change.

Pathohistological study of the ganglion plexuses of the sigmoid colon in patients with chronic slow-transit constipation

The morphological study of the resected sections of the colon obtained at the S.P. Fedorov Department of Faculty Surgery of S.M. Kirov Military Medical Academy, as a result of surgical treatment of patients with severe chronic slow-transit constipation, included an assessment of the changes in the structures of ganglion plexuses. Three cases were considered (women, aged 3740 years). Various degrees of pathological changes were detected in the ganglion plexuses (Auerbach and Meissner) of the sigmoid colon from patients with chronic slow-transit constipation using Nissls toluidine blue staining. In all cases, reactive, dystrophic, severe degenerative-necrotic changes of ganglion cells, as well as the details of their death, were described in detail. Along with pathological changes in nerve cells in the myenteric nerve plexus and gliosis, features of neuronglial relationships were described, and the death of ganglion cells in the human colon with the active participation of specialized astrocyte-like glial cells was also established for the first time. In the third case, a pattern of pronounced dysplasia and dysgangliogenesis was revealed in the myenteric ganglion plexus of the sigmoid colon, and the presence of diffuse lymphmonocytic infiltrates was noted in the circular muscle layer. Pathological changes in the enteral nervous system in chronic slow-transit constipation reflect neuropathy, which can serve as the main cause of impaired intestinal functions and of some symptoms.