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The first two objectives were to establish which stimulation parameters of kilohertz frequency alternating current (KHFAC) neuromodulation influence the effectiveness of pudendal nerve block and its safety. The third aim was to determine whether KHFAC neuromodulation of the pudendal nerve can relax the pelvic musculature, including the anal sphincter. Simulation experiments were conducted to establish which parameters can be adjusted to improve the effectiveness and safety of the nerve block. The outcome measures were block threshold (measure of effectiveness) and block threshold charge per phase (measure of safety). In vivo, the pudendal nerves in 11 male and 2 female anesthetized Sprague Dawley rats were stimulated in the range of 10 Hz to 40 kHz, and the effect on anal pressure was measured. The simulations showed that block threshold and block threshold charge per phase depend on waveform, interphase delay, electrode-to-axon distance, interpolar distance, and electrode array orientation. In vivo, the average anal pressure during unilateral KHFAC stimulation was significantly lower than the average peak anal pressure during low-frequency stimulation (p < 0.001). Stimulation with 20 kHz and 40 kHz (square wave, 10 V amplitude, 50% duty cycle, no interphase delay) induced the largest anal pressure decrease during both unilateral and bilateral stimulation. However, no statistically significant differences were detected between the different frequencies. This study showed that waveform, interphase delay and the alignment of the electrode along the nerve affect the effectiveness and safety of KHFAC stimulation. Additionally, we showed that KHFAC neuromodulation of the pudendal nerves with an electrode array effectively reduces anal pressure in rats.

Background: Acanthopanax senticosus (Rupr. et Maxim.) is commonly used in Traditional Chinese Medicine. Syringin is a major ingredient of phenolic glycoside in Acanthopanax senticosus. Objective: This study was performed to investigate whether Syringin could protect high glucose-induced bone marrow mesenchymal stem cells (BMSCs) injury, cell senescence, and osteoporosis by inhibiting JAK2/STAT3 signaling. Methods: BMSCs isolated from both the tibia and femur of mice were induced for osteogenesis. The cell senescence was induced using the high glucose medium. The cells were treated with 10 and 100 μmol/l Syringin. Immunohistochemistry staining was performed to determine the β-galactosidase (SA-β-gal) levels in differentially treated BMSCs. MTT assay and flow cytometry analysis were also performed to assess cell viability and cell cycle. The level of ROS in cells with different treatment was measured by using flow cytometry with DCF-DA staining. Calcium deposition and mineralized matrices were detected with alizarin red and ALP staining, respectively. Osteogenesis related genes OCN, ALP, Runx2, and BMP-2 were detected by RT-PCR. Levels of senescence-related proteins including p53 and p21, as well as JAK2, p-JAK2, STAT3, and p-STAT3 were detected by Western blot analysis. Results: Syringin treatment reversed the phenotypes of senescence caused by high glucose in BMSCs, including the arrest of G0/G1 cell cycle, enhanced SA-β-gal activity, and impaired cell growth. Syringin also decreased the elevated ROS production and the levels of p53, p21, and JAK2/STAT3 signaling activation. In addition, Syringin also enhanced the osteogenic potential determined by ARS and ALP staining, as well as increasing OCN, ALP, Runx2, and BMP-2 expressions. Conclusion: Syringin protects high glucose-induced BMSC injury, cell senescence, and osteoporosis by inhibiting JAK2/STAT3 signaling.

Objective: This research investigated the effects of different hemodialysis modalities combined with low-calcium dialysate (LCD) on mineral metabolism and vascular calcification (VC) in maintenance hemodialysis (MHD) patients with chronic kidney disease (CKD). Methods: General data were collected from 192 cases of MHD patients, who were divided into 4 groups according to the randomized numerical table. Each group was given LCD treatment, and conventional hemodialysis (HD), high-flux HD (HFHD), hemodiafiltration (HDF), and HD + hemoperfusion (HP) were performed, respectively. The patients were dialyzed 3 times per week for 4 h each time, and each group was treated for 6 months. Fasting venous blood was collected. Serum interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and high-sensitive C-reactive protein (hs-CRP) levels were measured by ELISA, calcium (Ca2+), phosphorus (P), Ca2+-P product, serum creatinine (SCr), blood urea nitrogen (BUN), β2 microglobulin (β2-MG), and intact parathyroid hormone (iPTH) were measured by chemiluminescence immunoassay, serum alkaline phosphatase (ALP) was determined by turbidimetric assay, and 25-hydroxyvitamin D (25(OH)D) was measured by autoradiographic immunoassay. To assess the extent of calcification in the iliac artery and abdominal aorta, a multilayer spiral CT device was employed for abdominal scans. Results: Serum IL-6, hs-CRP, TNF-α, Ca2+, P, Ca2+-P product, SCr, BUN, β2-MG, iPTH, and ALP levels decreased, while 25(OH)D levels increased in the four groups after treatment. The most pronounced effect on the reduction of IL-6, hs-CRP, TNF-α, Ca2+, P, Ca2+-P product, SCr, BUN, β2-MG, iPTH, and ALP was in the HD + HP group, followed by the HDF and HFHD groups, and then by the HD group. The rate of VC in the HDF, HFHD, and HD + HP groups was lower than that in the HD group, and the rate in the HD + HP group was lower than that in the HDF and HFHD groups. Conclusion: The combination of HD + HP and LCD in treating CKD with MHD is effective, evidently rectifying disruptions in serum Ca2+ and P metabolism, enhancing kidney function, lessening the body's inflammatory response, and lessening VC.

The single nucleotide polymorphism (SNP) A118G (rs1799971) in the Mu Opioid Receptor 1 (OPRM1) gene is associated with significant variations in analgesic doses and adverse effects of opioids. The A118G OPRM1 allele distributions vary significantly between different populations worldwide. The study aimed to assess the allele frequency and genotype distribution of OPRM1 A118G SNP in Saudis. This cross-sectional study included 124 healthy Saudis (62 males and 62 females) visiting the King Abdulaziz University Hospital in Jeddah, Saudi Arabia. The Oragene®-DISCOVER (OGR-600) kits were used to collect saliva samples from the participants. Polymerase chain reaction-restriction fragment length polymorphism was utilized to assess the SNP. Among the tested population, 79.03% (95% C.I. 70.81-85.82) were homozygous wild-type A118A, 16.13% (95% C.I. 10.14-23.80) were heterozygous A118G, and 4.84% (95% C.I. 1.80-10.23) were homozygous mutant G118G. OPRM1 A118G polymorphism allele frequencies were 87% (95% C.I. 79.89-92.44) and 13% (95% C.I. 7.56-20.11) for the 118A and 118G alleles, respectively. A higher frequency of the OPRM1 118G allele was present in females, 21% (95% C.I. 11.66-33.17) compared to males, 5% (95% C.I. 1.01-13.50). Relative to other Asian countries, the Saudi population showed a low prevalence of the OPRM1 A118G polymorphism, with a higher frequency of the 118G allele in females. Our research will contribute to the existing knowledge on the prevalence of OPRM1 A118G polymorphism, which could be considered for the personalized prescribing of opioid analgesics.

Introduction: False aneurysms in the thoracic aorta are dangerous complications that can occur after cardiac surgery. They often result in high mortality rates. These aneurysms are caused by damage to all layers of the aortic wall. This study aimed to pinpoint the area of the experimental specimen (native vessel, anastomosis, or prosthetic graft) with the greatest deformation, to determine whether a false aneurysm is likely to develop in the anastomotic portion. Methods: We conducted the inflation-extension test by performing eight cycles ranging from 0 to 20. The pressure sampling frequency was 100 Hz, and each cycle lasted approximately 34 seconds, resulting in a loading frequency of 0.03 Hz. During the experiment, each camera captured 3,000 frames. Based on the data collected, we evaluated and compared the loading stages of cycle 1 and cycle 8. Results and discussion: During loading, the native vessel experienced a dominant deformation of approximately 7% in the circumferential direction. The prosthetic graft, which had a longitudinal construction, deformed by approximately 8% in the axial direction. The prosthetic graft, on the other hand, only experienced a deformation of up to 1.5% in the circumferential direction, which was about 5 times smaller than the deformation of the native vessel. The anastomosis area was very stiff and showed minimal deformation. Additionally, there was little difference in the mechanical response between the first C1 and the eighth C8 cycle. Conclusion: Based on the available evidence, it can be inferred that aortic false aneurysms are more likely to form just behind the suture lines in the native aorta, which is more elastic compared to stiff sections of anastomosis and prosthetic graft. Numerous pulsations of the native vessel will likely cause the impairment of the aorta at the margin of the anastomosis. This will lead to disruption of the aortic wall and false aneurysm formation in the native vessel near the area of anastomosis.

In this study, the therapeutic efficacy of bergenin was examined against high-fat diet (HFD) induced type 2 diabetes in C57BL/6J mice. These mice were fed continuously HFD for 16 weeks and subjected to intragastric administration of various doses (10, 20 and 40 mg/kg body weight (BW)) of bergenin daily for subsequent 8 weeks. Bergenin supplementation to HFD-fed mice lower body weight gain, plasma glucose and insulin in diabetic mice. It further restored the alterations of biochemical parameters to near normal levels in diabetic mice. As compared to other two doses of 10 mg and 20 mg, bergenin 40 mg/kg BW were showed significant protective effect on the biochemical parameter studied. Consequently, it improved insulin-dependent glucose transport in the liver through translocation and activation of glucose transporter protein 2 (GLUT 2) in phosphatidylinositol 3-kinase (PI3K)/phosphorylated protein kinase B (AKT) dependent pathway. These findings provided evidence to exhibit that bergenin could improve liver tissue hyperglycemia, insulin sensitivity increase glucose uptake and shows hepatoprotective. Hence it might be used in the management of obesity-associated type 2 diabetes mellitus.

Due to its aggressive nature and low survival rate, esophageal cancer is one of the deadliest cancer. While the intestinal microbiome significantly influences human health and disease. This research aimed to investigate and characterize the relative abundance of intestinal bacterial composition in esophageal cancer patients. The fecal samples were collected from esophageal cancer patients (n = 15) and healthy volunteers (n = 10). The PCR-DGGE was carried out by focusing on the V3 region of the 16S rRNA gene, and qPCR was performed for Bacteroides vulgatus, Escherichia coli, Bifidobacterium, Clostridium leptum and Lactobacillus. High-throughput sequencing of the 16S rRNA gene targeting the V3+V4 region was performed on 20 randomly selected samples. PCR-DGGE and High-throughput diversity results showed a significant alteration of gut bacterial composition between the experimental and control groups, which indicates the gut microbial dysbiosis in esophageal cancer patients. At the phylum level, there was significant enrichment of Bacteroidetes, while a non-significant decrease of Firmicutes in the experimental group. At family statistics, a significantly higher level of Bacteroidaceae and Enterobacteriaceae, while a significantly lower abundance of Prevotellaceae and Veillonellaceae were observed. There was a significantly high prevalence of genera Bacteroides, Escherichia-Shigella, while a significantly lower abundance of Prevotella_9 and Dialister in the experimental group as compared to the control group. Furthermore, the species analysis also showed significantly raised level of Bacteroides vulgatus and Escherichia coli in the experimental group. These findings revealed a significant gut microbial dysbiosis in esophageal cancer patients. So, the current study can be used for the understanding of esophageal cancer treatment, disease pathway, mechanism, and probiotic development.

We have extracted and characterized Phasa fish (Setipinna phasa) oil for the first time to evaluate the anti-obesity and related anti-inflammatory effects on obese mice. Inbred male albino BALB/c mice were segregated into three categories: control (C), Obese control group (OC), and Phasa fish oil treated group (TX). To establish the potentiality of Setipinna phasa oil for its anti-obesity and anti-inflammatory properties, it was extracted and characterized using GC-MS method. To evaluate the anti-obesity effect, different parameters were considered, such as body weight, lipid composition, obesity, and obesity associated inflammation. The physicochemical characteristics of Phasa fish oil revealed that the oil quality was good because acid value, peroxide value, p-anisidine value, Totox value, refractive index, and saponification value were within the standard value range. The GC-MS study explored the presence of fatty acids beneficial to health such as Hexadec-9-enoic acid; Octadec-11-enoic acid; EPA, DHA, Methyl Linolenate, etc. The application of Setipinna phasa oil on the treated mice group acutely lowered body weight and serum lipid profile compared to the obese group. In connection with this, leptin, FAS, and pro-inflammatory cytokines TNF-α genes expression were downregulated in the treated group compared to the obese group. The Phasa oil treated group had an elevated expression of PPAR-α, adiponectin, LPL gene, and anti-inflammatory markers IL-10 and IL-1Ra compared to the obese group. This study suggests that Phasa fish oil, enriched with essential fatty acid, might be used as an anti-obesity and anti-inflammatory supplement.

Aim: This study aimed to investigate the phytochemical composition of Psychotria montana extract (PME) and evaluate its inhibitory effects on MCF7 breast cancer cells. Methods: The chemical composition of PME was analyzed using UPLC-QToF-MS. The effects of PME on cell proliferation were evaluated using the MTT assay. Flow cytometry was used for cell cycle and apoptosis analysis. The effects of PME on the transcription of cell cycle control genes were assessed using real-time PCR. Results: UPLC-QToF-MS analysis revealed major compounds of PME, including terpenoids and flavonoids, with the potential to inhibit proliferation, migration, and induce apoptosis in MCF7 cancer cells. PME effectively suppressed MCF7 cell proliferation under 2D culture, with a low IC50 value of 34.7 µg/ml. PME also hindered cell migration (p < 0.01) and reduced spheroid number (p < 0.001) and size (p < 0.001) in serum-free 3D culture. Apoptosis analysis via nuclear staining with DAPI and flow cytometry revealed an increase in the number of apoptotic cells after PME treatment (p < 0.001). Additionally, the PME induced cell cycle arrest at the G0/G1 phase (p < 0.05). PME altered the expression of cell cycle control genes (cyclins and CDKs) as well as cancer suppressor genes including p16, p27, and p53 at the transcriptional level (mRNA). The results of molecular docking suggest that the compounds present in PME exhibit a high binding affinity for CDK3, CDK4, CDK6, and CDK8 proteins, which are essential regulators of the cell cycle. Conclusion: Psychotria montana has the potential to inhibit cancer cells by inducing apoptosis and halting the cell cycle of MCF7 breast cancer cells

Atrial fibrillation (AF) is a common arrhythmia encountered in clinical practice, characterized by myocardial fibrosis and atrial remodeling as its primary pathological features, and associated with significantly high mortality and disability rates. Currently, there are no specific pharmacological treatments for AF, and traditional anti-arrhythmic drugs have not achieved the desired efficacy, often resulting in a high incidence of adverse drug reactions. Thus, there is an urgent need for the development of novel anti-AF medications. Berberine, the main active component of Coptis chinensis, has been shown to have antiarrhythmic and anti-heart failure effects. However, its potential to improve atrial fibrosis and remodeling resulting from AF remains largely unexplored. In this study, we used a rapid atrial pacing (RAP) procedure to establish a rabbit model of AF associated with atrial fibrosis. Our objective was to assess the inhibitory effects of berberine on myocardial fibrosis, evaluate its impact on atrial remodeling, and investigate its underlying molecular mechanisms. Our findings indicate that berberine reduces left atrial weight and the area of myocardial fibrosis, inhibits the expression of α-SMA protein in atrial tissue, and decreases the levels of inflammation and oxidative stress. In addition, berberine effectively inhibits atrial remodeling, which may contribute to the prevention of AF. Through transcriptomics, molecular docking, and molecular dynamics simulations, we have tentatively confirmed that berberine may activate the AMPK-PPARα signaling pathway by directly binding to AMPK and PPARα, thereby improving atrial fibrillation.

Introduction: The human nasal cavity and paranasal sinuses host a complex and dynamic microbiome which has a crucial role in mucosal immunity. A comprehensive profile of the healthy sinonasal microbiome remains limited. The purpose of our study was to characterize the healthy sinonasal microbiome in adults using 16S rRNA long-read sequencing to enable species-level resolution, and to assess its associations with demographical and clinical factors such as smoking, allergy history, and olfactory function. Study design: We performed a prospective, single-centre study analysing middle meatus samples from 27 healthy individuals undergoing septoplasty in the age range from 21 to 57 years, excluding those with antibiotic and corticosteroid use and those with signs of acute or chronic rhinosinusitis. Results: A high interindividual variability in the composition of healthy sinonasal microbiome was observed. At the phylum level, it was dominated by Firmicutes (48.96%), Actinobacteria (34.83%), and Proteobacteria (13.85%), while Firmicutes and Actinobacteria were consistently present in all samples. At the genus level, Staphylococcus spp. (32.32%), Cutibacterium (28.04%), and Corynebacterium (4.66%) were most abundant. We observed trend level correlations between phyla and some clinical factors (e.g., smoking and olfactory dysfunction) and selected phyla. However, none remained significant after false discovery rate (FDR) correction across taxa. Conclusion: The study proposes Staphylococcus spp., Corynebacterium spp., and Cutibacterium spp. to be a core taxa in the healthy sinonasal microbiome. Amid the interindividual diversity in our cohort, there was evidence of a stable core microbiome potentially influenced by environmental and host factors. Our findings suggest a baseline reference for distinguishing a dysbiosis in upper respiratory disease.

Introduction: ORBEYE^TM exoscope offers superior visualization with clearer imaging compared to standard methods and supports narrow band imaging (NBI). ORBEYE^TM provides better visualization of tissue structures, thus increasing the accuracy of the surgical procedure. The systematic use of ORBEYE^TM in transoral exoscopic laryngeal surgery (TOLES) is rarely documented. This study evaluates the sensitivity and specificity of TOLES using ORBEYE^TM with white light and NBI modalities. Methods: Between 8/2021 and 8/2024, 84 patients underwent TOLES using ORBEYE^TM with white light and NBI modes. All surgeries were performed in a specialized setting with consistent preoperative and intraoperative imaging protocols. Results: TOLES using ORBEYE^TM was successfully performed in all 84 patients. A statistically significant dependence (p < 0.001) was observed between preoperative/perioperative (TOLES) findings and definitive histology for both white light and NBI modalities. ORBEYE^TM facilitated detailed imaging of tissue structures and allowed targeted biopsies. Conclusion: TOLES with perioperative NBI/white light endoscopy achieved a high correlation between pre-histopathological diagnoses and final histopathological results. ORBEYE^TM exoscope provides three-dimensional 4K resolution, superior imaging, and improved ergonomics for the surgeon, reducing workload and increasing efficiency. It delivered more efficient surgical team collaboration and experience sharing. The integration of NBI into the TOLES system facilitated accurate navigation and targeting of the biopsy, helping to establish correct definitive diagnosis. TOLES showed more accurate scoring of perioperative NBI findings.

Embryonic stem (ES) cells are pluripotent cells widely used in cell therapy and tissue engineering. However, the broader clinical applications of ES cells are limited by their genomic instability and karyotypic abnormalities. Thus, understanding the mechanisms underlying ES cell karyotypic abnormalities is critical to optimizing their clinical use. In this study, we focused on proliferating human and mouse ES cells undergoing multipolar divisions. Specifically, we analyzed the frequency and outcomes of such divisions using a combination of time-lapse microscopy and cell tracking. This revealed that cells resulting from multipolar divisions were not only viable, but they also frequently underwent subsequent cell divisions. Our novel data also showed that in human and mouse ES cells, multipolar spindles allowed more robust escape from chromosome segregation control mechanisms than bipolar spindles. Considering the frequency of multipolar divisions in proliferating ES cells, it is conceivable that cell division errors underlie ES cell karyotypic instability.

Myo-inositol (MI), present in a variety of foods, is essential in several important processes of cell physiology. In this study, we explored the protective effects of MI against hyperglycemia and dyslipidemia in db/db mice, a typical animal model of type 2 diabetes mellitus (T2DM). MI supplement effectively suppressed the high plasma glucose and insulin levels and markedly relieved the insulin resistance (IR) in the db/db mice, comparable to metformin's effects. In MIN6 pancreatic β cells, MI also restrained the upsurge of insulin secretion stimulated by high-concentration glucose but had no impact on the promoted cell proliferation. Moreover, MI abated the enhanced plasma triglyceride and total cholesterol levels in the db/db mice. Notably, the lipid droplet formation of mesenchymal stem cells (MSCs) from db/db mice was significantly diminished after the treatment of MI, indicating that MI could effectively inhibit the differentiation of db/db mouse MSCs into adipocytes. However, MI regretfully failed to control obesity in db/db mice. This work proved that MI significantly helped db/db mice's metabolic disorders, indicating that MI has potential as an effective adjunctive treatment for hyperglycemia and dyslipidemia in T2DM patients.

Background: The current obstructive sleep apnea (OSA) diagnostic uses polysomnography or limited polygraphy and requires specialized personnel and technical equipment. Glycoprotein biomarkers and microRNAs are being explored as a possible new method for screening. We aimed to evaluate whether certain biomarkers and microRNA, previously identified as related to OSA, could be influenced by factors such as gender, age, and obesity level in patients with OSA. Methods: In this retrospective analytical study, patients with moderate to severe OSA (n = 130) were compared with the control group. Serum levels of selected biomarkers and microRNA were taken from both groups. The group of OSA patients was then stratified by gender, obesity level, and age to see the possible influence of those variables on biomarker levels. Results: Levels of all studied biomarkers - C-reactive protein (CRP), high-sensitivity troponin I (hsTnI), pentraxin-3 (PTX-3), and microRNA-499 were significantly higher in patients with OSA compared to the control group. In the OSA group only hsTnI showed a statistically significant relationship with gender. Levels of CRP and hsTnI showed a significant dependence on the level of obesity. Dependency on age was proven for hsTnI. CRP, PTX-3, and microRNA-499 did not have any statistically significant relationship with age. Conclusion: We found that serum levels of pentraxin-3 and microRNA-499 in patients with moderate to severe obstructive sleep apnoea are independent of gender, obesity, and age. CRP was affected by the level of obesity and hsTnI was influenced by all 3 variables. We consider these findings important for further research of OSA biomarkers.

This study investigates the potential relationship between exposure to polycyclic aromatic hydrocarbons (PAHs), specifically monohydroxylated metabolites (OH-PAHs), in urine, and the prevalence of respiratory diseases in 2-year-old children residing in two locations within the Czech Republic - České Budějovice (control location) and the historically contaminated mining district of Most. Despite current air quality and lifestyle similarities between the two cities, our research aims to uncover potential long-term health effects, building upon previous data indicating distinctive patterns in the Most population. A total of 248 urine samples were analysed for the presence of 11 OH-PAHs. Employing liquid-liquid extraction with ethyl acetate and clean-up through dispersive solid-phase extraction, instrumental analysis was conducted using ultra-high performance liquid chromatography coupled with tandem mass spectrometry. The incidence of respiratory diseases was assessed through questionnaires administered by paediatricians. The concentrations of OH-PAHs were elevated in urine samples from 2-year-olds in Most compared to those from České Budějovice. The incidence of respiratory diseases showed statistically significant higher levels of OH-PAHs in children from Most, together with a higher incidence of influenza. This association underlines the impact of environmental PAH exposure on children's respiratory health. It suggests that elevated urinary OH-PAH levels indicate an increased risk of developing respiratory diseases in the affected population. Further studies are needed to clarify the possible long-term health effects and to contribute to sound public health strategies.

Background: COVID-19 is a viral disease notorious for frequent worldwide outbreaks. It is difficult to control, thereby resulting in overload of the healthcare system. A possible solution to prevent overcrowding is rapid triage of patients, which makes it possible to focus care on the high-risk patients and minimize the impact of crowding on patient prognosis. Methods: The triage algorithm assessed self-sufficiency, oximetry, systolic blood pressure, and the Glasgow coma scale. Compliance with the triage protocol was defined as fulfillment of all protocol steps, including assignment of the correct level of care. Triage was considered successful if there was no change in the scope of care (e.g., unscheduled hospital admission, transfer to different level of care) or if there was unexpected death within 48 hours. Results: A total of 929 patients were enrolled in the study. Triage criteria were fulfilled in 825 (88.8%) patients. Within 48 hours, unscheduled hospital admission, transfer to different level of care, or unexpected death occurred in 56 (6.0%), 6 (0.6%), and 5 (0.5%) patients, respectively. The risk of unscheduled hospital admission or transfer to different level of care was significantly increased if triage criteria were not fulfilled [13.1% vs. 76.1%, RR 5.8 (3.8-8.3), p < 0.001; 0.5% vs. 5.2%, RR 11.4 (2.3-57.7), p = 0.036, respectively]. Conclusion: The proposed algorithm for triage of patients with proven COVID-19 is a simple, fast, and reliable tool for rapid sorting for outpatient treatment, hospitalization on a standard ward, or assignment to an intensive care unit.

In 2020, there were numerous cases in Kazakhstan with clinical symptoms of COVID-19 but negative PCR results in nasopharyngeal and oropharyngeal swabs. The diagnosis was confirmed clinically and by CT scans (computed tomography). The problem with such negative PCR results for SARS-CoV-2 infection confirmation still exists and indicates the need to confirm the diagnosis in the bronchoalveolar lavage in such cases. There is also a lack of information about confirmation of SARS-CoV-2 infection in deceased patients. In this study, various tissue materials, including lungs, bronchi, and trachea, were examined from eight patients who died, presumably from SARS-CoV-2 infection, between 2020 and 2022. Naso/oropharyngeal swabs taken from these patients in hospitals tested PCR negative for SARS-CoV-2. This study presents a modified RNA isolation method based on a comparison of the most used methods for RNA isolation in laboratories: QIAamp Viral RNA Mini Kit and TRIzol-based method. This modified nucleic acid extraction protocol can be used to confirm SARS-CoV-2 infection by RT-qPCR in the tissues of deceased patients in disputed cases. RT-qPCR with RNA of SARS-CoV-2 re-extracted with such method from post-mortem tissues that were stored at -80 °C for more than 32 months still demonstrated high-yielding positive results.

Background: Metabolic syndrome is a significant pro-inflammatory and pro-coagulant condition. The clinical association of adiponectin, a mainly antidiabetogenic molecule, and its interaction with platelets and platelet indices remains insufficiently investigated. Objective: The aim of our study was to investigate the association of adiponectin with platelets and platelet indices in patients with metabolic syndrome. Methods: The investigation was conducted as a cross-sectional study involving 113 subjects: 63 patients with the diagnosis of metabolic syndrome, and 50 healthy controls - with clear inclusion and exclusion criteria. The group of patients with metabolic syndrome was divided into two subgroups according to the platelet/high-density lipoprotein (HDL) ratio. Results: The subgroup with a higher platelet/HDL ratio was prediabetic. In the same subgroup of patients, a positive correlation between the adiponectin and mean platelet volume (MPV) was seen, while linear regression (95% CI) confirmed the association. Conclusion: Considering that MPV is the index that indicates average platelet volume and activity, we believe this association with adiponectin can represent a protective compensatory response in patients with metabolic syndrome and prediabetes. Our results provide a basis for a more precise selection of patients in whom the future therapeutic application of recombinant adiponectin would be most effective.

As a common complication of diabetes mellitus (DM), diabetic cardiomyopathy (DCM) is considered to be one of the major causes of mortality and morbidity. The therapeutic effects of naringenin have been verified in the treatment of various human diseases. However, the application of naringenin in the treatment of DCM still has not been reported. In this study, human AC16 cardiac cells were treated with normal d-glucose and high d-glucose (HG). After transfection with miR-30d-5p inhibitor, Cell Counting Kit-8 (CCK-8) method was used to measure cell viability. Hoechst 33258 staining was performed to observe the morphological changes of nucleus. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the activity of caspase-3. Cell apoptosis was detected by Annexin V-FITC/propidium iodide (PI) staining. Levels of light chain 3 (LC3) including LC3-I and LC3-II as well as nucleoporin p62 (P62) were detected by Western blot. We found that Naringenin treatment increased the reduced cell variability caused by HG treatment. Naringenin also increased expression level of miR-30d-5p in human AC16 cardiac cells after HG treatment. Treatment with miR-30d-5p inhibitor reduced the effect of miR-30d-5p in increasing cell variability and reducing cell apoptosis. Naringenin treatment reduced the increased levels of LC-I, LC-II and P62, but miR-30d-5p inhibitor reduced those changes. Therefore we concluded that naringenin could alleviate HG-induced injuries through the upregulation of microRNA-30d-5p level in human AC16 cardiac cells.

Background: Femoral posterior hip dislocation with associated femoral head fractures (Pipkin fractures) are rare high-energy injuries. Published treatment modalities involve conservative treatment, head fragment resection, open reduction and internal fixation, and total hip replacement. The experience with mini-invasive screw osteosynthesis of these fractures is the main focus of our study. Methods: Seven Pipkin fractures (five Pipkin II and two Pipkin I) in six patients were treated by closed reduction of hip dislocation, followed by minimal invasive lag screw osteosynthesis. Cancellous screw(s) were inserted from the incision on the lateral hip through the femoral neck to the reduced fracture fragment. In all patients, postoperative CT was performed to check the quality of surgery. Active physiotherapy with immediate toe-touch weight bearing was the routine postoperative protocol. In all patients, radiological and clinical results were evaluated with the Thompson Epstein, Merle d'Aubigne and Postel score, and Harris hip score. Results: All fractures united, and all femoral heads survived. Infectious complications were not observed, and no secondary surgery was needed. After an average follow-up of 18.4 months, the average Merle d'Aubigne and Postel score was 17.7 points, while the mean Harris hip score reached 98.1 points. The majority of patients achieved an excellent Thompson-Epstein clinical and radiological outcome. All patients returned to their original occupation. Conclusions: Mini-invasive screw osteosynthesis can be used for the treatment of Pipkin type I-II femoral head fractures. Successful reduction of hip dislocation and head fracture is necessary for using this technique. Long-term follow-up is necessary to confirm this technique.

Background: Both angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) are known to be effective in managing cardiovascular diseases, but more evidence supports the use of an ACEI. This study investigated the difference in cardiovascular disease incidence between relatively low-compliance ACEIs and high-compliance ARBs in the clinical setting. Methods: Patients who were first prescribed ACEIs or ARBs at two tertiary university hospitals in Korea were observed in this retrospective cohort study for the incidence of heart failure, angina, acute myocardial infarction, cerebrovascular disease, ischemic heart disease, and major adverse cardiovascular events for 5 years after the first prescription. Additionally, 5-year Kaplan-Meier survival curves were used based on the presence or absence of statins. Results: Overall, 2,945 and 9,189 patients were prescribed ACEIs and ARBs, respectively. When compared to ACEIs, the incidence of heart failure decreased by 52% in those taking ARBs (HR [95% CI] = 0.48 [0.39-0.60], P < 0.001), and the incidence of cerebrovascular disease increased by 62% (HR [95% CI] = 1.62 [1.26-2.07], P < 0.001). The incidence of ischemic heart disease (P = 0.223) and major adverse cardiovascular events (P = 0.374) did not differ significantly between the two groups. Conclusions: ARBs were not inferior to ACEIs in relation to reducing the incidence of cardiocerebrovascular disease in the clinical setting; however, there were slight differences for each disease. The greatest strength of real-world evidence is that it allows the follow-up of specific drug use, including drug compliance. Large-scale studies on the effects of relatively low-compliance ACEIs and high-compliance ARBs on cardiocerebrovascular disease are warranted in the future.

Increasing data has confirmed the potential anticancer properties of Dendrobium, a traditional Chinese herb. However, most anticancer compositions from the plant of Dendrobium were usually extracted by high polar solvent, while weak polar compositions with excellent anticancer activity remained largely unexplored. In this study, the differences between ether extract and ethanol extract of Dendrobium nobile Lindl. on chemical components and anticancer activities were investigated, as well as the anticancer mechanisms among different extracts. The results demonstrated that the ether extract exhibited a stronger anticancer effect than ethanol extract, and its anticancer effect was mainly due to weak polar compounds rather than polysaccharides and alkaloids. Quantitative proteomics suggested that the ether extract significantly stimulated the over-expression of immature proteins, the endoplasmic reticulum stress and unfolded protein response were subsequently induced, the intracellular reactive oxygen species level was seriously elevated, and oxidative stress occurred in the meanwhile. Eventually, autophagy and apoptosis were activated to cause cell death. Our findings demonstrate that the ether extract of D. nobile is a potential candidate for anticancer drug development, and that future research on anticancer drugs derived from medicinal plants should also concentrate on weak polar compounds.

Skin cancer has high rates of mortality and therapeutic failure. In this study, to develop a multi-agent strategy for skin cancer management, the selective cytotoxicity of several alkaloid fractions and pure alkaloids isolated from Amaryllidaceae species was evaluated in melanoma cells. In addition, UVB-stimulated keratinocytes (HaCaT) were exposed to seven alkaloid fractions characterized by GC-MS, and the production of intracellular reactive oxygen species (ROS) and IL-6, were measured to evaluate their photoprotection effects. The Eucharis caucana (bulb) alkaloid fraction (20 μg/ml) had a clear effect on the viability of melanoma cells, reducing it by 45.7% without affecting healthy keratinocytes. This alkaloid fraction and tazettine (both at 2.5 μg/ml) suppressed UVB-induced ROS production by 31.6% and 29.4%, respectively. The highest anti-inflammatory potential was shown by the Zephyranthes carinata (bulb) alkaloid fraction (10 μg/ml), which reduced IL-6 production by 90.8%. According to the chemometric analysis, lycoramine and tazettine had a photoprotective effect on the UVB-exposed HaCaT cells, attenuating the production of ROS and IL-6. These results suggest that Amaryllidaceae alkaloids have photoprotective and therapeutic potential in skin cancer management, especially at low concentrations.

Background: Atrial fibrillation is common in patients with structural heart disease who are undergoing cardiac surgery. Surgical CryoMaze has been shown to be an effective treatment in several trials, but success rates have varied considerably, between 47-95%. The sequential hybrid approach, combining surgical CryoMaze followed by radiofrequency catheter ablation, can achieve high freedom from atrial arrhythmias. However, in patients with concomitant surgical atrial fibrillation treatment, data comparing the hybrid approach to CryoMaze alone are lacking. Methods: The SurHyb study was designed as a prospective, open-label, multicentre randomized trial. Patients with non-paroxysmal atrial fibrillation who were scheduled for coronary artery bypass grafting or valve repair/replacement were randomized to either surgical CryoMaze alone or surgical CryoMaze followed by radiofrequency catheter ablation 3 months post-surgery. The primary outcome measure was arrhythmia-free survival without class I or III antiarrhythmic drugs, which has been evaluated using implantable cardiac monitors. Conclusions: This is the first randomized study that compares concomitant surgical CryoMaze alone with the staged hybrid surgical CryoMaze followed by catheter ablation, in patients with non-paroxysmal atrial fibrillation using rigorous rhythm monitoring. The results may contribute to the optimization of the treatment in patients undergoing concomitant CryoMaze for atrial fibrillation.

Background: Xanthine oxidase (XO) generates reactive oxygen species during uric acid production. Therefore, XO inhibitors, which suppress oxidative stress, may effectively treat non-alcoholic steatohepatitis (NASH) and atherosclerosis via uric acid reduction. In this study, we examined the antioxidant effect of the XO inhibitor febuxostat on NASH and atherosclerosis in stroke-prone spontaneously hypertensive 5 (SHRSP5/Dmcr) rats. Methods: SHRSP5/Dmcr rats were divided into three groups: SHRSP5/Dmcr + high-fat and high-cholesterol (HFC) diet [control group, n = 5], SHRSP5/Dmcr + HFC diet + 10% fructose (40 ml/day) [fructose group, n = 5], and SHRSP5/Dmcr + HFC diet + 10% fructose (40 ml/day) + febuxostat (1.0 mg/kg/day) [febuxostat group, n = 5]. Glucose and insulin resistance, blood biochemistry, histopathological staining, endothelial function, and oxidative stress markers were evaluated. Results: Febuxostat reduced the plasma uric acid levels. Oxidative stress-related genes were downregulated, whereas antioxidant factor-related genes were upregulated in the febuxostat group compared with those in the fructose group. Febuxostat also ameliorated inflammation, fibrosis, and lipid accumulation in the liver. Mesenteric lipid deposition decreased in the arteries, and aortic endothelial function improved in the febuxostat group. Conclusions: Overall, the XO inhibitor febuxostat exerted protective effects against NASH and atherosclerosis in SHRSP5/Dmcr rats.

Introduction: Narrow band imaging (NBI) is an endoscopic imaging method intended for the diagnosis of mucosal lesions of the larynx that are not visible in white-light endoscopy, but are typical of pre-tumor and tumor lesions of the larynx. The purpose of the study: To compare preoperative/perioperative white light endoscopy and NBI endoscopy with the results of histopathological examinations in pre-tumor and tumor lesions of the larynx. Methods: A prospective study, over a period of five years (5/2018-5/2023), included 87 patients with laryngeal lesions aged 24-80 years. We evaluated preoperative/ perioperative white light and NBI endoscopy, established a working prehistological diagnosis, and compared this with the definitive histopathological results of laryngeal biopsies. Results: In relation to the definitive histology score, a statistically significant correlation was found between the evaluation of the finding and the definitive histology for preoperative and perioperative white light endoscopy and NBI endoscopy (p < 0.001). Both methods showed higher precision when used perioperatively. Conclusion: NBI endoscopy is an optical method that allows us to improve the diagnosis of laryngeal lesions, perform a controlled perioperative biopsy, and refine the surgical scope. The NBI endoscopy is a suitable method for the diagnosis of early cancerous lesions of the larynx. The use of preoperative/perioperative NBI endoscopy allowed us to achieve a high level of agreement correlation (p < 0.001) between the prehistological working diagnosis and the final histopathological result. The NBI method proves its application in the diagnosis of pre-tumor and tumor lesions of the larynx.

Aim: We investigated the antimicrobial and anticancer properties of an ethanol crude extract of Red Sea brown alga (Hormophysa cuneiformis) from Egypt. Methods: Extraction was achieved by mixing 100 g of sample powder with absolute ethanol, incubating at 37 °C overnight in a shaking incubator, and then collecting the extract. The extract's antimicrobial activity was tested using a well diffusion assay against the tested pathogens (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Candida albicans) in comparison to commercial antibiotics. Anticancer activity was assessed using MTT assay on MCF-7, HepG-2, and HEP-2 cell lines. The anticancer mechanism of action against the HepG-2 cell line was investigated using cell cycle analysis, Annexin V, and antioxidant enzymes, in addition to transmission electron microscopy. Results: GC-MS phytoconstituent profile of the extract was dominant with fatty acids. A broad antimicrobial effect against all the pathogenic isolates of E. coli, S. aureus, B. subtitles, and C. albicans was demonstrated, especially at the high concentration in comparison to commercial antibiotics. The extract could inhibit the growth of the tested cell lines. We observed the most significant effect on HepG-2 cells, and the concentration of the extract played a role in the level of inhibition (IC50 of 44.6 ± 0.6 µg/ml). The extract had negligible effects on Vero normal cell lines at the lower concentration, with slight toxicity (90.8% viability) at the highest concentration (500 µg/ml). At this same concentration, the extract caused 80-92% inhibition of the cancer cell lines. The extract appears to have demonstrated promising effects on cancer cells. It induces programmed cell death (apoptosis), arrests the cell cycle, and affects the oxidative/antioxidant balance within the cells, potentially leading to the suppression or elimination of cancer cells. These findings are encouraging and may have implications for cancer treatment or further research in this area. More action of extract was seen against bacteria than fungi, with a wide antibacterial impact against all of the tested isolates, notably at the high concentration in comparison to conventional antibiotics. Conclusion: According to the findings, H. cuneiformis may be a valuable source of chemicals that are both antimicrobial and anticancer.

Bacillary dysentery (shigellosis) continues to cause havoc worldwide, with a high infectivity rate. It causes bloody diarrhea, and around 99% of bacillary dysentery cases occur in developing countries. The objective of this study is to develop a polyherbal formulation with the scientific rationale in treating infectious bacillary dysentery disease. The anti-bacterial activity, the minimum inhibitory concentration of the formulation against bacillary dysentery, causing microbes like Shigella flexneri (MTCC 1457), Escherichia coli (MTCC 1687), and Salmonella enterica (MTCC 98), was analysed by well-diffusion method and broth dilution method, respectively. The biofilm inhibition activity was determined on 96 well polystyrene plates and anti-quorum sensing activity by Chromobacterium violaceum CV026. The cytotoxicity was examined by acute oral toxicity. Excreta and organ bacterial load were analyzed by serial dilution method. The formulation efficacy was determined by analyzing the blood sample of rats. The antimicrobial efficacy of the developed formulation was calculated by measuring the zone of inhibition which was found to be 24 mm, 25 mm, and 25 mm, and the MIC values of 1.5 mg/ml, 1.5 mg/ml, and 2.0 mg/ml against S. flexneri, S. enterica, E. coli, respectively. The results show that the polyherbal formulation significantly reduced biofilm formation and has anti-quorum sensing activity. The formulation also effectively decreases the bacterial load and increases the K+, Na+, and Ca++ ions in animals treated with the formulation. The developed formulation was found to be non-toxic and effective against bacillary dysentery; thus, it can be used for treating bacillary dysentery and related complications.

Background: Interferon-gamma (IFN-γ) is a chief proinflammatory cytokine with a significant role in the immune response against viral infections. Today there is increasing evidence about the association between individual genetic polymorphisms and cytokines in predicting HBV infection susceptibility. Aim: This study aimed to investigate the association between IFN-γ gene polymorphisms and susceptibility to hepatitis B viral infection (HBV), and the impact of these genetic polymorphisms on IFN-γ production. IFN-γ (+874A/T, rs2430561, and +2109A/G, rs1861494) was genotyped by single-stranded polymorphism-polymerase chain reaction (SSP-PCR) in 126 Egyptians with chronic HBV infection and in 100 healthy control subjects. The plasma levels of IFN-γ were measured by Enzyme-linked immunosorbent assay (ELISA). Results: Compared to the control subjects there was a slight increase in +874TT genotype frequency in HBV patients. However, no statistical significance in IFN-γ (+874A/T and +2109A/G) genotype/allele distribution was demonstrated, indicating the lack of association between these SNPs and susceptibility to HBV infection. In +2109A/G, only AG genotype was observed with a complete abrogation of GG and AA genotypes. Haplotypes between different loci on selected genes showed insignificant changes in their frequency in patients and control subjects. HBV patients had a significantly higher level of IFN-γ (P < 0.001) compared to controls. The maximum significant increase in IFN-γ production was observed in subjects harboring the +874TA genotype. Conclusions: As no association could be characterized between the polymorphism in IFN-γ (+874A/T and +2109A/G) and susceptibility to chronic HBV infection, our data support the concept that IFN-γ gene polymorphisms are not predictors of HBV susceptibility in this segment of the Egyptian population.