Paraffin Sections Research Articles

Development of a Rapid Diagnostic Method for Sporotrichosis.

Sporotrichosis, a prevalent deep fungal infection in clinical settings, currently lacks rapid and accurate diagnostic methodologies. This study explores a novel rapid diagnosis method for sporotrichosis by combining FTA cards and nested PCR with fungal fluorescence staining. The study involved skin lesion tissues from 26 patients diagnosed with sporotrichosis (Experimental Group). The Positive Control Group consisted of fungal suspensions from clinical strains of Sporothrix, while the Negative Control Group included fungal solutions of other fungi, namely Trichophyton rubrum, Trichophyton mentagrophyte, and Candida albicans. DNA was extracted from the slurry of skin lesions in the Experimental Group and from fungal suspensions in the Control Group using FTA cards, followed by nested PCR amplification. Subsequently, nested PCR amplification was performed. Histopathological examinations, including HE and fluorescence staining, were conducted on paraffin sections prepared from skin lesion tissues in the Experimental Group. Among the 26 clinical skin lesion tissues in the Experimental Group, 8 cases showed a specific positive band upon nested PCR amplification, resulting in a positive rate of 30.8%. In the Control Group, the fungal solution of the clinical strain of Sporothrix showed a specific positive band upon nested PCR amplification, while all other fungi Negative Control Group tested negative. Histopathological examination revealed granulomatous inflammatory changes in most samples after HE staining. Fluorescence staining detected spores in 17 cases, resulting in a detection rate of 65.4% (17/26). The combination of FTA cards with nested PCR method proved to be simple and rapid but demonstrated a relatively low positive rate. Fungal fluorescence staining significantly improved the sensitivity of detecting sporotrichosis in histopathological examinations, thereby improving the speed and efficiency of diagnosis.

Studies on Distant Hybridization Compatibility between the Azalea (Rhododendron × hybridum hort.) and the Rhododendron decorum Franch. Native to China

Rhododendron resources are abundant in China, and hybridization breeding technology is the primary method for cultivating Rhododendron varieties. In order to optimize the utilization of wild Rhododendron resources for distant hybridization, this study took three horticultural varieties of Rhododendron subgenus Tsutsusi and the Rhododendron decorum Franch. of the subgenus Hymenanthes as the research objects, and the cross-compatibility between subgenera was analyzed from the aspects of pollen tube growth and ovary and seed development. At the same time, the statistics of ovary swelling rate and fruit bearing rate, numbers of capsule seeds, 1000 seed weight of hybrid seeds, germination rate, green seedling rate, and other indexes were analyzed to comprehensively evaluate hybrid fertility. The results showed that there was obvious pre-fertilization and post-fertilization barriers existing in the hybridization of Rhododendron × hybridum hort. and R. decorum. During the growth of pollen tubes, a large amount of callus appeared, which led to the entanglement, distortion, and abnormal development of the pollen tubes; only part of the pollen tubes entered into the ovary or ovule. The pre-fertilization barriers can be overcome by early pollination and delayed pollination. According to the observation of the ovary paraffin section, 45 d after pollination, the seed was shriveled and developed abnormally. The comprehensive evaluation of hybrid fertility showed that ‘Sima’ × R. decorum was fertile, ‘Yin Taohong’ × R. decorum was a weakly fertile, and ‘Little Taohong’ × R. decorum was sterile. This study provided a scientific basis for intergeneric hybridization breeding between the subgenus Tsutsusi and the subgenus Hymenanthes.

First record of Phoronopsis (Phoronida) from Japan, with a description of Phoronopsiscalifornica Hilton, 1930 from Okinawa.

Phylum Phoronida currently contains two genera, Phoronis Wright, 1856 and Phoronopsis Gilchrist, 1907, with approximately thirteen speices. Phoronida is distributed worldwide, ranging from northern Europe to southern New Zealand and also from intertidal to 400 m depths. From Japanese waters, four species of Phoronis have been reported, viz. Phoronisijimai Oka, 1897, Phoronisaustralis Haswell, 1883, Phoronispsammophila Cori, 1889 and Phoronisemigi Hirose et al., 2014. We describe the morphology of Phoronopsiscalifornica Hilton, 1930, based on five specimens collected at a sandy-mud habitat in Nago Bay and Oura Bay, Okinawa, Japan. We examined the internal anatomy by serial paraffin sections. We also examined the genetic distance of Japanese specimens from the other phoronid sequences in GenBank. This is the first record of Phoronopsis from Japanese waters and the fifth species record of phoronids in addition to the previously recorded four species of Phoronis.

Fatty acid profiles unveiled: gene expression in Yanbian yellow cattle adipose tissues offers new insights into lipid metabolism

Abstract. Objectives. The objectives of this study were twofold: to analyze the composition and content of fatty acids in various adipose tissues (including kidney, abdominal, subcutaneous, and omental) of Yanbian yellow cattle and to observe the morphology of adipocytes within these tissues and to assess the level of expression of specific genes – kinase insert domain receptor (KDR), apolipoprotein L domain containing 1 (APOLD1), stearoyl-CoA desaturase 1 (SCD1), secreted frizzled-related protein 4 (SFRP4), fatty-acid-binding protein 5 (FABP5), and sterol carrier protein-2 (SCP2) – in different adipose tissues (kidney, abdominal, posterior belly, ribeye, prothorax, striploin, upper brain, and neck) of Yanbian yellow cattle. Method. Castrated Yanbian yellow cattle, 24 months old, with identical genetic backgrounds and raised under the same breeding management conditions, were selected. The fatty acid composition and content were assessed using gas chromatography, while the size and diameter of adipocytes were analyzed via paraffin sectioning. The level of expression was determined using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Results. In total, 16 distinct fatty acids were identified in abdominal adipose tissue. Additionally, henicosanoic acid (C21:0) and behenic acid (C22:0) were detected exclusively in subcutaneous adipose tissue. Caprylic acid (C8:0) was found in both kidney and omental adipose tissues. The size of individual adipocytes in kidney adipose tissue was notably larger compared to the adipocytes in the other three regions (p<0.05). Regarding gene expression, APOLD1 exhibits its highest expression in striploin adipose tissues (p<0.05), while SCD1 shows its peak expression in prothorax adipose tissues (p<0.05). Moreover, both FABP5 and SCP2 demonstrate their highest level of expression in prothorax adipose tissue (p<0.05). Furthermore, the level of expression of KDR and SFRP4 across these seven adipose tissue regions exhibits significant differences (p<0.05). Conclusion. In conclusion, Yanbian yellow cattle exhibit variations in both the composition and content of fatty acids across different adipose tissue depots, including the kidney, abdominal, subcutaneous, and omental regions. Moreover, adipocytes display distinct morphological differences across these tissue types. Furthermore, the level of expression of KDR, APOLD1, SCD1, SFRP4, FABP5, and SCP2 varies significantly among adipose tissues located in the kidney, abdominal, posterior belly, ribeye, prothorax, striploin, upper brain, and neck regions.

Gastric Precancerous Lesions: From Progenitor Cell and Microsatellite Instability to Clinical Interpretation of Gastric Cancer Risk

Aim: to evaluate the possibility of the MMR-system status, microsatellite instability (MSI) usage in the differential diagnosis of gastric mucosa dysplasia, determination of the gastric adenocarcinoma development risk. Material and methods. The study included gastric mucosa specimens of 75 patients: 25 with high-grade dysplasia, 25 with low-grade dysplasia, 25 were indefinite for dysplasia. Gastrobiopsy specimens were examined histologically, immunohistochemically using mouse monoclonal antibodies (Diagnostic BioSystems, USA) to the MMR system proteins: MLH-1 (clone G168-15, dilution 1:50), MSH2 (clone DBM15.82, dilution 1:100), MSH6 (clone 44, dilution 1:50), PMS2 (clone A16-4, ready to use). MSI was studied with multiplex PCR evaluation of DNA microsatellites (NR-21, NR-24, NR-27, BAT-25, BAT-26) from paraffin sections, their analysis with capillary electrophoresis. The obtained data were processed with the Statistica 10.0 (StatSoft, USA), presented using descriptive, analytical statistics. VOSviewer (1.6.20) was used to visualize the bibliometric analysis. Results. MMR-deficient cases were found in low (2.8 %) and high-grade (2.8 %) dysplasia with the immunohistochemical evaluation of MMR-system proteins in gastric mucosa specimens. In all indefinite for dysplasia cases MMR-system proteins remained unaffected. Three MSI-positive cases (6.5 %) were detected by PCR with two low-grade dysplasia, one high-grade dysplasia cases. All identified cases were also immunohistochemically MSI-positive. Conclusion. Determination of MSI can be used as an auxiliary study within a panel of biomarkers aimed to support the decision-making of a pathologist in the alternative of “indefinite for dysplasia” or “definite dysplasia — obligate precancer”.

Establishing of 3D-FISH on frozen section and its applying in chromosome territories analysis in Populus trichocarpa.

Fluorescence in situ hybridization with frozen sections of root tips showed difference of chromosome territories distribution between autosome and sex-chromosome homologous pairs in Populus trichocarpa. The spatial organization of chromatin within the interphase nucleus and the interactions between chromosome territories (CTs) are essential for various biologic processes. Three-dimensional fluorescence in situ hybridization (3D-FISH) is a powerful tool for analyzing CTs, but its application in plants is limited. In this study, we established a 3D-FISH technique using frozen sections of Populus trichocarpa root tips, which was an improvement over the use of paraffin sections and enabled us to acquire good FISH signals. Using chromosome-specific oligo probes, we were able to analyze CTs in interphase nuclei in three dimensions. The distribution of chromosome pairs 17 and 19 in the 3D-preserved nuclei of P. trichocarpa root tip cells were analyzed and showed that the autosome pair 17 associated more often than sex chromosome 19. This research lays a foundation for further study of the spatial position of chromosomes in the nucleus and the relationship between gene expression and spatial localization of chromosomes in poplar.

The value and significance of nucleolar organizer region proteins as markers of malignancy in breast cancer patients.

To assess argyrophilic nucleolar organizer regions (AgNORs) in 60 patients with primary breast carcinoma and evaluated their association with clinical prognostic parameters of breast cancer. Argyrophilic nucleolar organizer regions were stained in paraffin sections of the tissues using Ploton's silver method. For each sample, the number of AgNORs within the nuclei of 100 tumor cells was counted. The average number of AgNORs per nucleus was calculated, and the results were expressed as mean. The number of AgNORs was significantly higher in breast invasive ductal carcinoma (6.6) compared to benign breast tumors (fewer than 2.0). However, differences in AgNOR counts across different age groups were not statistically significant. This study suggests that AgNOR counts could be used as a potential procedure for estimating proliferation characteristics in histopathological sections of benign and malignant breast lesions. Argyrophilic nucleolar organizer region counts may also be valuable for identifying high-risk patients and indicating tumor aggressiveness. A larger study with an increased sample size that incorporates both AgNOR numbers and Ki67 scores for assessing cell kinetics is needed to confirm our findings.

Identification and Biological Characterization of Green Alga on Oil-Tea Camellia Leaves

Between 2023 and 2024, a type of green alga was observed for the first time settling on Oil-tea Camellia leaves and branches in the eastern Oil-tea Camellia planting area of Hainan Island, forming a layer of gray-green moss with a rough surface that seriously interfered with the leaves’ normal photosynthesis. To further research the issue, this study used the plant photosynthesis measurement system and the paraffin sectioning technique to compare and analyze the changes in photosynthetic characteristics and anatomical structure of healthy and green algal-covered Oil-tea Camellia leaves. At the same time, the algal strain was effectively separated and purified using the plate delineation method, and its species classification was determined by combining morphological observation and molecular identification based on SSU-ITS sequences. The results of the study demonstrated that the coating of green alga facilitated the lignification of the leaf’s epidermal cell walls. After being covered by the green alga, the intercellular CO2 concentration (Ci) increased significantly by 21.5%, while the net photosynthetic rate (Pn), transpiration rate (Tr), and stomatal conductance (Gs) all significantly decreased by 72.8%, 30.4%, and 24.9%, respectively. More specifically, the green alga that covers the leaves of Oil-tea Camellia was identified as Desmodesmus armatus of Chlorophyta. Notably, the green alga had a long growth cycle, did not undergo a decline phase within one month, had an optimal growth pH of 11.0, and could flourish in excessively alkaline conditions. In conclusion, this study not only reported for the first time the phenomena of D. armatus infesting Oil-tea Camellia leave, but also showed its unique physiological and ecological properties, providing a foundation for future research on relevant prevention and control approaches.

Metabolomics reveals a key role of salicylic acid in embryo abortion underlying interspecific hybridization between Hydrangea macrophylla and H. arborescens.

Embryo abortion at the heart-shaped stage is the main reason for the failure of interspecific hybridization of hydrangea, and salicylic acid plays a key role during embryo abortion. Difficulties in obtaining seeds from interspecific hybridization between Hydrangea macrophylla and H. arborescens had severely restricted the process of breeding new hydrangea varieties. To clarify the cause of reproductive barriers, an interspecific hybridization was made between H. macrophylla 'Endless Summer' (female parent) and H. arborescens 'Annabelle' (male parent). The results showed that both parents' floral organs developed normally, 'Annabelle' had high pollen viability (84.83% at 8h after incubation), and the pollen tube could enter into the ovule of 'Endless Summer' at 72h after pollination. Therefore, the pre-fertilization barrier was not the main reason for the failure of interspecific hybridization. However, observation of the embryo development by paraffin sections showed that the embryo was aborted at the heart-shaped stage. In addition, salicylic acid (SA) content was significantly higher (fourfold, P < 0.01) at 21 days after pollination (DAP) as compared to that of 17 DAP, which means SA may be closely correlated with embryo development. A total of 957 metabolites were detected, among which 78 were significantly different. During the embryo abortion, phenylpropanoids and polyketides were significantly down-regulated, while organic oxygen compounds were significantly up-regulated. Further analysis indicated that the metabolic pathway was enriched in the shikimic acid biosynthesis pathway, which suggests that more SA was synthesized. Taken together, it can be reasonably speculated that SA plays a key role leading to embryo abortion underlying the interspecific hybridization between Hydrangea macrophylla and H. arborescens. The result is helpful to direct the breeding of hydrangea through distant hybridization.

Characterization of Banana Crowns: Microscopic Observations and Macroscopic Cutting Experiments

Banana crowns’ intricate vascular systems facilitate nutrient transport for fruit growth and provide mechanical support. Analyzing vascular bundle morphology facilitates understanding of its influence on the banana de-handing process. In this study, we employed X-ray Computed Tomography (CT) scanning and microscopic observation of paraffin sections to characterize the morphological traits of the banana crown’s vascular tissue system and reconstructed its 3D vascular tissue system throughout the banana bunch. Based on the internal tissue characteristics and external morphology, the banana crown is categorized into three regions: the central stalk–crown transition region (CSCTR), the crown expansion region (CER), and the crown–finger transition region (CFTR). Cutting experiments indicated that variations in the cutting strength and specific cutting energy across positions within the banana bunch were insignificant but significantly distinct among the three regions. Specifically, the CER showed a 19.7% reduction in cutting strength and a 15.5% decrease in energy consumption compared to the other regions. This was due to the unique cross-distribution of fibers within the CER, which were primarily parallel to the cutting blade, significantly reducing cutting forces and energy consumption, making the CER the optimal region for cutting. The orientation of vascular bundles relative to the blade is key to optimizing plant cutting mechanics.

Histological and histochemical study of effect of dietary fibre in motility of stomach in male mice.

To investigate the relationship between dietary fibres and muscularis externa layer. The experimental study was conducted at the Anatomy Department of the College of Medicine, Al- Nahrain University, Iraq, from November 2020 to April 2021, and comprised adult healthy males. They were divided randomly into experimental group A and control group B. Group A was fed high-fibre diet, and group B was fed standard pellet diet. The tissue samples were harvested at day 30 post-surgery. The stomach samplings were placed in 10% neutral formalin for 24 hours to obtain paraffin sections for routine histological and immunohistochemical staining. The protein expression in stomach's smooth muscle of each group was detected by immunohistochemical staining. Data was analysed using SPSS 24. Of the 20 mice, 10(50%) were in each of the two groups. Group A exhibited significant weight-gain compared to group B (p≤0.01). There was significant reduction in the thickness of the muscularis layer in group A compared to group B (p≤0.01). Anoctamin 1 expression in group A was significantly lower than that in group B (p≤0.01). The stress induced by an unstable fibre diet significantly affected the stomach by decreasing the muscularis layer thickness and Anoctamin 1 expression in interstitial cells of Cajal.

Effects of protein diet on expression of Anoctamin1 of Cajal cell in the nervous plexus of the stomach in male mice.

To determine the association between a protein-rich diet and the expression of anoctamin 1 in Interstitial cells of Cajal within the muscle layer of the stomach wall in male mice. The experimental study was conducted at the Anatomy Department of the College of Medicine, Al- Nahrain University, Iraq, from November 2020 to April 2021, and comprised male adult healthy male mice. They were divided randomly into two equal groups. Group A was fed a high protein diet, while control group B was fed a standard pellet diet. The tissue samples were harvested at day 30 post-surgery. The stomach samplings were placed in 10% neutral formalin for 24 hours to obtain paraffin sections for routine histological and immunohistochemical staining. The protein expression in stomach smooth muscle of each group was detected by immunohistochemical staining. Data was analysed using SPSS 24. Of the 20 mice, 10(50%) were in each of the two groups. Group A exhibited significant weight-gain compared to group B (p≤ 0.05). There was significant elevation in muscle wall thickness in group A, compared to group B (p≤0.05). Tunica muscularis of the stomach in group A significantly thickened compared to group B (p≤0.05). The expression of anoctamin 1 was significantly more intense in group A compared to group B (p≤0.01). Unbalanced food had a significant impact on the stomach, affecting the thickness of the muscularis layer and the expression of anoctamin 1 in cajal cells in the muscularis externa.

Remodeling of the structural components of the capsule and glomerular zone of the adrenal glands cortex of white rats under the influence of a complex of food additives at the late terms of the experimental study

With food additives, we can add flavour to food, extend its shelf life, and make it look good. However, even small amounts of these additives can lead to diseases of various organs and systems. The aim of our work is to establish metrics and morphological changes in the structure of the capsule and the glomerular zone of the cortex of the adrenal glands of rats at the later stages of the experimental study under the conditions of consumption of a complex of food additives: monosodium glutamate, sodium nitrite and Ponceau 4R. The control group of rats consumed oral drinking water and saline. The rats of the experimental groups were orally administered once a day with a 10 % solution of sodium nitrite (E250), sodium glutamate (E621) or Ponceau 4R. The dosages of food additives were two times lower than the permissible norm in food. The rats were withdrawn from the experiment after 8, 12 and 16 weeks. The adrenal glands were removed for the study. Paraffin sections were stained with hematoxylin and eosin, and semi-thin sections with toluidine blue, and examined using a light microscope Biorex with a digital microphoto attachment DSM 900. Ultrathin sections were examined in a TEM-125 K electron microscope. For morphometric analysis, the structure of the capsule and the glomerular zone of the cortex of the adrenal glands of rats at the later stages of the experimental study was studied. The study results were analysed using the software package “InStat”. At the late stages of the experimental study, the adrenal capsule underwent irreversible changes. The average thickness of the adrenal connective tissue capsule decreased by the end of the 16th week of the experimental research. Similarly, the average thickness of the cortex decreased with each subsequent week relative to the control values, indicating significant structural and functional disorders. The average thickness of the glomerular zone of the adrenal cortex periodically changed as a compensatory reaction in response to the administration of a complex of food additives to rats. The phenomena of nucleus pyknosis and sludge syndrome were detected, which indicated compensatory and adaptive processes at the microscopic level. The analysis of electron micrographs revealed destroyed granular endoplasmic reticulum cisternae and mitochondrial remnants. Thus, it has been established that using a complex of food additives (monosodium glutamate, sodium nitrite and Ponceau 4R) directly affects the adrenal glands of rats. This effect causes structural changes in the cortex and medulla, causing alteration and hypertrophy of cells in these areas. Additionally, hyperhydration of the connective tissue stroma and compensatory-restorative reactions aimed at transcription factors and internal mechanisms of nitric oxide formation were recorded.

Structural changes in the organs of the lymphoid system in terms of induced carcinogenesis

Colorectal cancer is the third most common cancer in the world. Despite recent therapeutic advances, it causes more than 500,000 deaths each year. The immune system plays a crucial role in protecting the body from cancer. However, cancer cells are able to evade immune detection and destruction. For example, they can downregulate antigen expression, produce immunosuppressive molecules, or recruit immune cells that perform regulatory or inhibitory functions. Understanding the complex interactions between the immune system and carcinogenesis is crucial for developing effective cancer treatments. The study aimed to determine the morphological changes in the thymus, spleen, and lymph nodes under N,N-dimethylhydrazine-induced carcinogenesis. The study was performed on 77 male outbred albino rats weighing 190-230 g, kept in standard vivarium conditions. Colon adenocarcinoma was modeled by administration of N,N-dimethylhydrazine hydrochloride for 30 weeks. To study the peculiarities of morphological and functional changes in lymphoid organs in the dynamics of colon tumor lesions, animals were withdrawn from the experiment every 30 days. Paraffin sections of the thymus, spleen, and lymph nodes 5-6 μm thick were made on a rotary microtome and stained with hematoxylin and eosin. The experimental investigation unveiled the character and extent of histological alterations within the lymphoid system organs under conditions of N,N-dimethylhydrazine-induced colon adenocarcinomatosis. As the experiment progressed, there was a noticeable escalation in the severity of detrimental and degenerative modifications observed in the thymus, spleen, and lymph nodes. These modifications were evident in the disruption of blood circulation within the examined organs, leading to vascular wall impairment and hemorrhaging, the disarray of morphofunctional elements and the development of fibrosis. Given the significant role played by the thymus, spleen, and lymph nodes in regulating carcinogenesis and maintaining immune balance, it is extremely important to delve into understanding the changes in their structure and function. Obtained results indicate that carcinogenesis is accompanied by pronounced morphological changes in the structural components of the lymphoid system organs, the degree of which increases in direct proportion to the duration of exposure to the oncogenic factor.

The morphological structure of prostate gland under the condition of experimental prostatopathy and after using of cholecalciferol in the different schemes of hypofertility correction

The impact of negative factors, stress, and modern living conditions damages men's health and leads to infertility. Prostatitis is often a cause of hypofertility. It is now shown that vitamin D may play a role in regulating the functioning of reproductive system organs. The aim of the study was to determine the effect of cholecalciferol on the histological structure of the prostate gland in rats with experimental prostatitis and after its application alone or in combination with a prostate protector. Experimental prostatitis was induced by cold intraoperative damage to the prostate gland. To correct prostatitis, vitamin D3 (cholecalciferol) was administered orally at a dose of 4000 IU. The prostate protector (Prostatilen, Pr) and its pharmaceutical composition, as well as vitamin D3, were administered rectally. Rats with modeled prostatitis were divided into groups: EP (cold experimental prostatitis without treatment); EP + seed oil (on the background of experimental prostatitis, animals received a solvent – apricot kernel oil); EP + vit D3 (per os) (on the background of experimental prostatitis, animals received vitamin D3); EP + Pr (rec) (on the background of experimental prostatitis, males were administered Prostatilen per rectum); EP + vit. D3 (per os) + Pr (rec) (on the background of experimental prostatitis, animals received vitamin D3 (per os) and Prostatilen gel (per rectum)); EP + (vit. D3 + Pr) (rec) (on the background of experimental prostatitis, rats were administered Prostatilen gel with vitamin D3 per rectum). Intact animals (Intact group) and sham-operated rats (Control group) were used as controls. Paraffin sections of the prostate gland were stained with hematoxylin, eosin, and Van Gieson's method. In addition to the review microscopy of the ventral lobes of the prostate gland and the isthmus between them, the power of histochemical reactions was measured, the severity of inflammation and fibrosis was assessed, the number of terminal sections of the prostatic glands with a visually unchanged state, with lumen expansion, and with wall destruction was counted, the longitudinal diameter of the acini and the height of the epithelial cells of the prostatic glands were measured. Statistical analysis of the results was performed using the standard software package "Statistica 6.0" with the use of Student's t-test and nonparametric analog of one-way analysis of variance – Kruskal-Wallis test, as well as Mann-Whitney test. It was found that rats with experimental prostatitis exhibit pronounced changes in the morphological structure of the prostate gland. The prostate-protective effect of vitamin D per os at a dose of 4000 IU was established, which reduced the manifestations of atrophic and destructive processes, signs of tissue inflammation, and coarsening of the prostate gland stroma. Signs of fibrosis development in the prostate gland in males of this group were reduced, and the number of destructive changes and the longitudinal diameter of the terminal sections of the prostatic glands of prostate ventral lobe in rats with experimental prostatitis were decreased. Thus, the addition of cholecalciferol to the basic therapy for infertility has a more pronounced corrective effect on the morphological structure of the prostate than the separate use of cholecalciferol and the prostate protector. Combining basic therapy with vitamin D enhances the prostate-protective properties of the latter and is promising for restoring reproductive function overall.

Morphometric Characteristics of Postmortem Changes in the Brain of Newborns Depending on the Duration of the Postmortem Period.

The morphometric characteristics of brain tissue were studied based on autopsy material from 49 deceased newborns divided into 7 groups based on the time after death. Samples were taken from the upper (frontal lobe) and lower (occipital lobe) regions relative to the supine position of the body. Paraffin sections were prepared from these samples and stained with hematoxylin and eosin. Histological preparations were analyzed using an image analysis system to determine the area of gliocyte nuclei, cross-cut blood vessels, and expanded area around them in the white matter of the brain. Based on these data, severity indices were calculated for both cellular (pericellular) and vascular (perivascular) expansion. The dynamics of changes in morphometric parameters within the range of cellular and vascular alterations on brain tissue specimens, which reflect the development of postmortem hypostasis and autolysis in this organ, can be used to estimate the duration of the postmortem interval. At the same time, these changes, when combined with other non-specific postmortem findings, should be distinguished from long-term pathological processes and diseases that may have occurred during life.

The Effects of STRA6 Regulation of the Circadian Rhythm on Choroidal Neovascularization.

This study aims to investigate the relationship among STRA6, circadian rhythm, and choroidal neovascularization (CNV) formation, as well as the regulatory mechanism of STRA6 in CNV under circadian rhythm disturbances. C57BL/6J male mice (aged 6 weeks) were randomly divided into control and jet lag groups (using a time shift method every 4days to disrupt the molecular clock's capacity to synchronize with a stable rhythm). A laser-induced CNV model was established in both the control and the jet lag group after 2 weeks of jet lag. The size of CNV lesions and vascular leakage were detected by morphological and imaging examination on the seventh day post laser. STRA6 was screened by full transcriptome sequencing. Bioinformatics analysis was conducted to assess the variation and association of STRA6 in the GSE29801 dataset. The effects of STRA6 were evaluated both in vivo and in vitro. The pathway mechanism was further elucidated and confirmed through immunofluorescence of paraffin sections and Western blotting. The disturbance of circadian rhythm promotes the formation of CNV. Patients with age-related macular degeneration (AMD) exhibited higher levels of STRA6 expression compared to the control group, and STRA6 was enriched in pathways related to angiogenesis. In addition, CLOCK and BMAL1, which are initiators that drive the circadian cycle, had regulatory effects on STRA6. Knocking down STRA6 reversed the promotion of CNV formation caused by circadian rhythm disturbance in vivo, and it also affected the proliferation, migration, and VEGF secretion of RPE cells without circadian rhythm in vitro, as well as impacting endothelial cells. Through activation of the JAK2/STAT3/VEGFA signaling pathway in unsynchronized RPE cells, STRA6 promotes CNV formation. This study suggests that STRA6 reduces CNV production by inhibiting JAK2/STAT3 phosphorylation after circadian rhythm disturbance. The results suggest that STRA6 may be a new direction for the treatment of AMD.

Ultrastructural and thermal analyses reveal novel insights into low-temperature survival mechanisms of hydrated seeds of Poaceae species from alpine regions

Global warming leads to snow cover loss in the alpine ecosystem, exposing seeds to extreme diurnal temperature fluctuations during the growing season. The risk of freezing increases as seeds have increased moisture content. Studying the survival mechanisms of seeds at low temperatures can help analyze changes in alpine meadow populations and target conservation efforts. Here, we used three species of Poaceae as a model to understand freezing stress.Fully imbibed Elymus dahuricus, Festuca elata, and Lolium multiflorum seeds were subjected to programmed cooling at fast and slow rates (-1.0/0.05°C/min) and then assessed for survival. Differential Scanning Calorimetry was used to analyze thermal transitions during cooling. HE-stained paraffin sections and a Transmission Electron Microscope were employed to observe internal morphology and ultrastructural changes.E. dahuricus seeds exhibited greater tolerance to low temperatures than those of the other two species, with an LT50 of approximately -20°C for both cooling rates and maintained relatively intact ultrastructure. The observed Low-temperature exotherm (LTE) correlated with seed survival, with viability decreasing extensively below LTE.Fast cooling caused fewer changes to seed morphology and ultrastructure than slow cooling, suggesting that the primary survival mechanism during fast cooling is freezing avoidance through supercooling. Seeds exhibited greater freeze tolerance under slow than fast cooling, primarily by migrating intracellular water to extracellular spaces where it froze, causing considerable damage to cell ultrastructure and forming apparent cavities in some seeds.

Pathology diagnosis of intraoperative frozen thyroid lesions assisted by deep learning

BackgroundThyroid cancer is a common thyroid malignancy. The majority of thyroid lesion needs intraoperative frozen pathology diagnosis, which provides important information for precision operation. As digital whole slide images (WSIs) develop, deep learning methods for histopathological classification of the thyroid gland (paraffin sections) have achieved outstanding results. Our current study is to clarify whether deep learning assists pathology diagnosis for intraoperative frozen thyroid lesions or not.MethodsWe propose an artificial intelligence-assisted diagnostic system for frozen thyroid lesions that applies prior knowledge in tandem with a dichotomous judgment of whether the lesion is cancerous or not and a quadratic judgment of the type of cancerous lesion to categorize the frozen thyroid lesions into five categories: papillary thyroid carcinoma, medullary thyroid carcinoma, anaplastic thyroid carcinoma, follicular thyroid tumor, and non-cancerous lesion. We obtained 4409 frozen digital pathology sections (WSI) of thyroid from the First Affiliated Hospital of Sun Yat-sen University (SYSUFH) to train and test the model, and the performance was validated by a six-fold cross validation, 101 papillary microcarcinoma sections of thyroid were used to validate the system’s sensitivity, and 1388 WSIs of thyroid were used for the evaluation of the external dataset. The deep learning models were compared in terms of several metrics such as accuracy, F1 score, recall, precision and AUC (Area Under Curve).ResultsWe developed the first deep learning-based frozen thyroid diagnostic classifier for histopathological WSI classification of papillary carcinoma, medullary carcinoma, follicular tumor, anaplastic carcinoma, and non-carcinoma lesion. On test slides, the system had an accuracy of 0.9459, a precision of 0.9475, and an AUC of 0.9955. In the papillary carcinoma test slides, the system was able to accurately predict even lesions as small as 2 mm in diameter. Tested with the acceleration component, the cut processing can be performed in 346.12 s and the visual inference prediction results can be obtained in 98.61 s, thus meeting the time requirements for intraoperative diagnosis. Our study employs a deep learning approach for high-precision classification of intraoperative frozen thyroid lesion distribution in the clinical setting, which has potential clinical implications for assisting pathologists and precision surgery of thyroid lesions.

Exploration of Genes Related to Intramuscular Fat Deposition in Xinjiang Brown Cattle.

The aim of this study was to investigate the differentially expressed genes associated with intramuscular fat deposition in the longissimus dorsi muscle of Xinjiang Brown Bulls. The longissimus dorsi muscles of 10 Xinjiang Brown Bulls were selected under the same feeding conditions. The intramuscular fat content of muscle samples was determined by the Soxhlet extraction method, for which 5 samples with high intramuscular fat content (HIMF group) and 5 samples with low intramuscular fat content (LIMF group) were selected. It was found that the intramuscular fat content of the HIMF group was 46.054% higher than that of the LIMF group. Muscle samples produced by paraffin sectioning were selected for morphological observation. It was found that the fat richness of the HIMF group was better than that of the LIMF group. Transcriptome sequencing technology was used to analyze the gene expression differences of longissimus dorsi muscle. Through in-depth analysis of the longissimus dorsi muscle by transcriptome sequencing technology, we screened a total of 165 differentially expressed genes. The results of Gene Ontology (GO) enrichment analysis showed that the differentially expressed genes in the two groups were mainly clustered in biological pathways related to carbohydrate metabolic processes, redox processes and oxidoreductase activities. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the differentially expressed genes were significantly clustered in 15 metabolic pathways, which mainly covered fatty acid metabolism (related to lipid metabolism and glucose metabolism), the pentose phosphate pathway, the Peroxisome Proliferator-Activated Receptor (PPAR) signaling pathway and other important metabolic processes. The three genes that were predominantly enriched in the glycolipid metabolic pathway by analysis were SCD5, CPT1C and FBP2, all of which directly or indirectly affect intramuscular fat deposition. In summary, the present study investigated the differences in gene expression between high and low intramuscular fat content in the longissimus dorsi muscle of Xinjiang Brown Bulls by transcriptome sequencing technology and revealed the related signaling pathways. Therefore, we hypothesized that SCD5, CPT1C and FBP2 were the key genes responsible for the significant differences in intramuscular fat content of the longissimus dorsi muscles in a population of Xinjiang Brown Bulls. We expect that these findings will provide fundamental support for subsequent studies exploring key genes affecting fat deposition characteristics in Xinjiang Brown Bulls.