Rat Blood Research Articles

Activity of Ethanol Extract of Kalangkala seed (Litseaangulata B.) as Nephroprotector and Hepatoprotector in Alloxan-induced Diabetic Rats

Kalangkala (Litsea angulata B) is a traditional medicine that contains phenolic compounds and flavonoids and has antioxidant activity. Phenolic and flavonoids are reported to protect liver and kidney function and maintain insulin levels in the body. The purpose of this study was to determine the effect of administration of ethanol extract of kalangkala seeds (EEBK) as nephroprotective and hepatoprotective agents in rats (Rattus norvegicus L) induced by alloxan. Kalangkala seed was extracted by ethanol and evaporated to get the concentrated extract. The 30 male Wistar rats were divided into 6 groups including normal group, negative group, metformin positive group and treated groups which treated by EEBK 100, 200 and 400 mg/kgBW. Alloxan was injected intraperitoneally, metformin and EEBK were given orally for 21 days. The blood of rats was taken through the orbital sinus on day 21. The results showed the total flavonoid content of EEBK was 6.27 mgQE/g. EEBK doses of 100, 200 and 400 mg/kg BW as well as metformin can reduce (p<0.05) creatinine levels, ureum and SGPT activity but only EEBK doses of 200 and 400 mg/kgBW (p<0.05) can reduce SGOT activity. The administration of EEBK doses of 100. 200 and 400 mg / kgBW was able to improve the histopathological picture of the liver. It can be concluded that EEBK has the potential as a nephroprotective and hepatoprotective in diabetic rats (Rattusnorvegicus L) induced by alloxan

Evaluation of the Level of Low-Density Lipoproteins in the Blood of Rats with Rhabdomyolytic Renal Failure of Various Severity Degrees and Water-Soluble C60 Fullerenes Action

Evaluation of the Level of Low-Density Lipoproteins in the Blood of Rats with Rhabdomyolytic Renal Failure of Various Severity Degrees and Water-Soluble C60 Fullerenes Action

Bushen Zhuyun Decoction Enhances Endometrial Receptivity via the IL-6/STAT3 Signaling Pathway in Rats

Background: Reproductive endocrine disorder can impair endometrial receptivity, preventing embryo implantation and increasing miscarriage risk. Impaired endometrial receptivity contributes significantly to female infertility. Inflammatory signaling pathways including the IL-6/STAT3 pathway help embryos implant. Therefore, it is crucial to explore the relationship between the IL-6/STAT3 signaling pathway and endometrial receptivity. Objective: To investigate the mechanism by which Bushen Zhuyun decoction (BSZY) enhances endometrial receptivity in rats through the IL-6/STAT3 signaling pathway. Methods: Mifepristone-induced poor endometrial receptivity models of female SD rats were established, followed by histopathological observation. ELISA was used to measure serum sex hormones and VEGF. Western blotting or IHC was used to measure steroid receptors, IGFBP1, and IL-6/STAT3 pathway activation in the uterus during each estrus cycle and early gestation of normal rats. The Treg/Th17 balance was assessed using flow cytometry. Results: Significant differences were found in the protein expressions of steroid receptors, IL6, STAT3, and p-STAT3 during each estrus cycle and early gestation of normal rats. The protein expressions of STAT3 and PR were strongly correlated with each other. BSZY notably improved uterine morphology increased the expression of implantation markers and raised the serum concentrations of sex hormones and VEGF. BSZY enhanced the expressions of IL-6 and its receptors and restored the expressions of STAT3 and p-STAT3 in the uterus of pregnant rats. In addition, BSZY effectively restored the Treg/Th17 balance in the peripheral blood of pregnant rats. Conclusion: BSZY enhances endometrial receptivity and promotes decidualization in SD rats via the IL-6/STAT3 signaling pathway.

Creation of a database of historical control of hematological indices in rat blood to interpret the results of sanitary and toxicological studies of active substances of pesticides

Introduction. Historical control data provide an understanding of the background variability (spontaneous changes) of the studied indices in the same species and line of animals, under similar conditions in the same laboratory, when assessing the toxic effects of repeated (manifold) administration of active substances of pesticides. The aim of the study was to create a database of historical control of hematological indices in rat blood used in chronic (twelve months) studies of active substances of pesticides of various chemical structures. Materials and methods. In accordance with the planned goal, a database of hematological blood indicces from control animals used in chronic (12 months) sanitary and toxicological studies of active substances of pesticides of various chemical structures has been created. The database contains information on 13 hematological indices in rat blood taken from control animals in dynamics after 1, 3, 6 and 12 months from the start of the study for the period 2017–2022. Studies of the active substances of pesticides were carried out in the biological testing laboratory of the Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing according to the manual P 1.2.3156–13. Results. The results of the conducted studies indicate that in dynamics after 1, 3, 6 and 12 months of the study, a statistically significant increase in hemoglobin levels was detected in the blood of male rats. With the age, the animals showed a tendency to increase the concentration of red blood cells and platelets. Limitations. This article publishes a database of historical control of hematological parameters in rat blood, data on biochemical indices of blood serum and physiological indices in male rats in similar studies will be presented in the following publications. Conclusion. The results obtained will be used to substantiate the maximum inactive doses (NOEL), permissible daily doses (DSD), hygienic standards of the studied compounds in food and environmental objects.

Pterostilbene Targets Hallmarks of Aging in the Gene Expression Landscape in Blood of Healthy Rats.

Polyphenols from the phytoestrogen group, including pterostilbene (PTS), are known for their antioxidant, anti-inflammatory, and anti-cancer effects. In recent reports, phytoestrogens attenuate age-related diseases; however, their pro-longevity effects in healthy models in mammals remain unknown. As longevity research demonstrates age-related transcriptomic signatures in human blood, the current study hypothesizes that phytoestrogen-supplemented diet may induce changes in gene expression that ultimately confer pro-longevity benefits. In the present study, RNA sequencing is conducted to determine transcriptome-wide changes in gene expression in whole blood of healthy rats consuming diets supplemented with phytoestrogens. Ortholog cell deconvolution is applied to analyze the omics data. The study discovered that PTS leads to changes in the gene expression landscape and PTS-target genes are associated with functions counteracting hallmarks of aging, including genomic instability, epigenetic alterations, compromised autophagy, mitochondrial dysfunction, deregulated nutrient sensing, altered intercellular interaction, and loss of proteostasis. These functions bridge together under anti-inflammatory effects through multiple pathways, including immunometabolism, where changes in cellular metabolism (e.g., ribosome biogenesis) impact the immune system. The findings provide a rationale for pre-clinical and clinical longevity studies and encourage investigations on PTS in maintaining cellular homeostasis, decelerating the process of aging, and improving conditions with chronic inflammation.

L-Arginine and Intermittent Hypoxia Are Stress-Limiting Factors in Male Wistar Rat Models.

The aim of this study was to evaluate the combined effects of L-arginine, intermittent hypoxia training (IHT), and acute stress on oxygen-dependent processes in rats, including mitochondrial oxidative phosphorylation, microsomal oxidation, and the intensity of lipoperoxidation processes. In addition, our study investigated how the modulatory effect of the NO synthase mechanism on the concentration of catecholamines (CA), such as adrenaline and noradrenaline, and their biosynthetic precursors (DOPA, dopamine) varies depending on the cholinergic (acetylcholine, Ach-acetylcholinesterase, AChE) status in rats. This study investigated the protective stress-limiting effects of L-arginine impact and IHT in the blood and liver of rats. The results showed that L-arginine promoted the maintenance of NAD-dependent oxidation in mitochondria, which was detrimental compared to succinate oxidation, and was accompanied by depletion of respiratory activity reserves under stress induced by high concentrations of CA. The interdependence of SC-dependent oxidation and the functional role of NAD-dependent substrate oxidation in the mitochondrial respiratory chain in stress conditions induced using inhibitors revealed the importance of the NO system. Administration of L-arginine during the IHT course prior to stress exposure increased the compensatory capacity of the organism. L-arginine increased the compensatory capacity of the sympathoadrenal system in stress-exposed rats. In the early stages of IHT, modulation of the CA concentration was observed with a concomitant increase in lipoperoxidation processes, and in the final stages of IHT, the CA concentrations increased, but there was also an inhibition of lipoperoxidation, which was particularly enhanced by the administration of L-arginine. The increase in blood concentrations of CA and ACh was accompanied by a decrease in AChE activity at different stages of adaptation to hypoxia induced by IHT (days 5, 10, and 14). Thus, the IHT method significantly mobilises the reserve capacity of oxygen-dependent processes through the system of CA, ACh-AChE mediated by nitric oxide.

Estimation of Postmorterm Interval by Using Myoglobin Concentration in Blood, Liver And Parotid Glands of Albino Rats

Objective: To determine the postmortem interval (PMI) by estimating myoglobin concentration in blood, liver and parotid gland of albino rats and to compare the postmortem level of myoglobin among blood, liver and parotid gland. Study Design: Quasi-Experimental study. Place and Duration of Study: Department of Forensic Medicine and Toxicology King Edward Medical University, Lahore in collaboration with the University of Veterinary and Animal Sciences, Lahore from 20 June 2021 to 19 June 2022. Methodology: A total of 72 rats were killed by cervical dislocation and postmortem myoglobin levels were checked in blood, liver and parotid gland at 8 different postmortem intervals (9 rats in each interval) to check its significance in relation to postmortem time. Results: There was a significant correlation between myoglobin concentration in blood, liver and parotid gland with Postmortem Interval with p value of < 0.001. The analysis indicated that the combination of blood, liver, and parotid myoglobin concentrations yielded the highest predictive accuracy (r = 0.982). The next best model included liver and parotid myoglobin concentrations (r = 0.980), while the model based solely on liver myoglobin concentration had the lowest predictive accuracy (r = 0.946). Conclusion: The myoglobin concentration in glandular tissues is a helpful parameter to determine PMI. As the postmortem interval increases the myoglobin concentration in the blood, liver and parotid gland also increases.

Upregulation of P-Glycoprotein and Breast Cancer Resistance Protein Activity in Newly Developed <i>in Vitro</i> Rat Blood–Brain Barrier Spheroids Using Advanced Glycation End-Products

Upregulation of P-Glycoprotein and Breast Cancer Resistance Protein Activity in Newly Developed <i>in Vitro</i> Rat Blood–Brain Barrier Spheroids Using Advanced Glycation End-Products

C60 Fullerene Reduces the Development of Post-Traumatic Dysfunction in Rat Soleus Muscle.

Traumatic skeletal muscle injury is a complex pathology caused by high-energy trauma to muscle tissue. Previously, a positive effect was established when C60 fullerene was administered against the background of muscle ischemia, mechanical muscle injury, and other muscle dysfunctions, which probably protected the muscle tissue from damage caused by oxidative stress. Using tensiometry and biochemical analysis, the biomechanical parameters of skeletal muscle contraction and biochemical indices of the blood of rats 15 days after traumatic injury of the soleus muscle caused by myocyte destruction by compression were studied. The intraperitoneal administration of C60 fullerene aqueous solution (C60FAS) in a daily dose of 1 mg/kg improved its contractile function by 28-40 ± 2% and the values of the investigated biochemical indices of the animals' blood by 15-34 ± 2% relative to the trauma group. The obtained results indicate the potential ability of C60 fullerenes, as powerful antioxidants, to reduce the development of post-traumatic dysfunction of the soleus muscle.

Transgenic rat with ubiquitous expression of angiotensin-(1-7)-producing fusion protein: a new tool to study the role of protective arm of the renin-angiotensin system in the pathophysiology of cardio-renal diseases.

The aim of the present study was to assess systemic circulatory and tissue activities of both the classical arm and of the alternative arm of the renin-angiotensin system (RAS) in a new transgenic rat line (TG7371) that expresses angiotensin-(1-7) (ANG 1-7)-producing fusion protein; the results were compared with the activities measured in control transgene-negative Hannover Sprague-Dawley (HanSD) rats. Plasma and tissue concentrations of angiotensin II (ANG II) and ANG 1-7, and kidney mRNA expressions of receptors responsible for biological actions of ANG II and ANG 1-7 [i.e. ANG II type 1 and type 2 (AT1 and AT2) and Mas receptors] were assessed in TG7371 transgene-positive and in HanSD rats. We found that male TG7371 transgene-positive rats exhibited significantly elevated plasma, kidney, heart and lung ANG 1-7 concentrations as compared with control male HanSD rats; by contrast, there was no significant difference in ANG II concentrations and no significant differences in mRNA expression of AT1, AT2 and Mas receptors. In addition, we found that in male TG7371 transgene-positive rats blood pressure was lower than in male HanSD rats. These data indicate that the balance between the classical arm and the alternative arm of the RAS was in male TGR7371 transgene-positive rats markedly shifted in favor of the latter. In conclusion, TG7371 transgene-positive rats represent a new powerful tool to study the long-term role of the alternative arm of the RAS in the pathophysiology and potentially in the treatment of cardio-renal diseases.

Aromatherapy was used to explore the sedative and hypnotic effects of Moringa seed essential oil on insomnia rats

AbstractMoringa is a type of plant that is used both for medicinal and food. Moringa seed (MS) are rich in volatile oil and have initially been employed to treat diseases of the nervous system. Insomnia, a prevalent neurological disorder, has led to this study's aim: to extract the essential oil from MS and analyze its potential to improve sleep. This study utilized petroleum ether for the thermal extraction of the essential oil from MS, which was then subjected to compositional analysis using Gas Chromatograph Mass Spectrometer (GC–MS). P‐chlorophenyl alanine (PCPA) was used to induce an insomnia model in Sprague–Dawley (SD) rats. Following the successful establishment of the model, the MS essential oil was administered at concentrations of 10%, 5%, and 2.5% to investigate its sedative and hypnotic effects. The efficacy of the MS essential oil was assessed by observing the general condition of rats in each group, conducting an open field test, a pentobarbital sodium righting test, and measuring the serum 5‐HT (5‐hydroxytryptamine) levels and hypothalamic GABA (γ‐aminobutyric acid) content. GC–MS analysis of the MS essential oil revealed a rich composition, including oleic acid, palmitoleic acid, stigmasterol, and γ‐stigmasterol, among other substances. Through the assessment of the rats' general condition, behavioral tests, and blood biochemical assays, it was inferred that MS essential oil aromatherapy can reduce the rat's locomotor activity, increase their interest in activity and exploration, enhance the serum 5‐HT levels, and elevate hypothalamic GABA content. Consequently, it can be concluded that MS essential oil has a sedative and hypnotic effect.

STUDY OF TOXICOLOGY OF THE ANTITUMOR DRUG COLCHIPRIT-NEO (K-20)

Objective: Colchiprit-neo was administered orally to rats at dosages of 30.0, 61.0, and 120 mg/kg for 30 days in order to assess its chronic toxicity. Methods: The effects on behavior, body weight, peripheral blood composition, kidney function, liver enzyme levels, and internal organ pathology were evaluated after rats were given Colchiprit-neo at the recommended dosages. To assess the reversibility of any possible harmful effects, a one-month recovery time was incorporated. Results: According to the study, rats' behavior, body weight, and blood composition were unaffected by repeated oral treatment of Colchiprit-neo. During therapy, the liver's alanine aminotransferase (ALT) level rose, but during the recovery phase, all tested values went back to normal. The 30 mg/kg dose did not cause any toxicity in the parenchymal organs, according to pathomorphological analysis. Novelty: The findings indicate that Colchiprit-neo is safe for long-term usage because it does not show substantial chronic toxicity at therapeutic levels. This work offers significant preclinical evidence in favor of its possible therapeutic use without endangering the major organ systems.

Dan-shen Yin attenuates myocardial fibrosis after myocardial infarction in rats: Molecular mechanism insights by integrated transcriptomics and network pharmacology analysis and experimental validation

Ethnopharmacological relevanceDan-shen Yin (DSY) originated from the “Shi Fang Ge Kuo” is a Chinese formula composed of three medicines: Salvia miltiorrhiza (Dan-shen), Santalum album L. (Tan-xiang) and Amomum villosum Lour. (Sha-ren). It has many years of clinical experience in the prevention of myocardial fibrosis (MF). However, the specific mechanism of DSY in prevent MF is not clear. Aim of the studyThis study aimed to assess the efficacy of DSY in the prevention of MF and reveal its underlying mechanism in a rat model of MF after myocardial infarction (MI) induced by ligation of the left anterior descending branch (LAD) of the coronary artery. Materials and methodsThe blood-entry components of DSY were analyzed by UHPLC-Q-TOF-MS/MS. LAD-ligated rats were used to assess the efficacy of DSY in the prevention of MF. Network pharmacology and transcriptomics analysis were used to predict possible target signaling pathways of DSY in MF. Echocardiography, immunohistochemistry and ELISA methods were used to evaluate the cardiac functions and biochemical changes of the rats. The mRNA expressions of target genes were measured by RT-qPCR. The proteins expressions, including Collagen I, Collagen III, α-smooth muscle actin (α-SMA), matrix metallopeptidase 2 (MMP 2), matrix metallopeptidase 9 (MMP 9), transforming growth factor-β (TGF-β), protein kinase B (AKT), phospho-AKT, extracellular regulated protein kinases (ERK), phospho-ERK, c-Jun N-terminal kinase (JNK), phospho-JNK, mothers against decapentaplegic protein (Smad3), and phospho-Smad3 were detected and quantified by Western Blot. ResultsUHPLC-Q-TOF-MS/MS analysis disclosed that 20 components within DSY could be absorbed into blood of rats. DSY improved myocardial injury in the myocardial tissue of LAD-ligated rats, as evidenced by the elevation of left ventricular ejection fraction and left ventricular fractional shortening, and the decrease of the serum CK-MB and LDH levels. Network pharmacology and transcriptomics predicted that DSY could interfere biological processes, such as extracellular matrix organization, focal adhesion and ECM-receptor interaction, and modulate TGF-β mediated signaling pathways, including PI3K/AKT, MAPK, and Smad3. Further study confirmed that DSY reduced MF, accompanied by reduced TGF-β, Collagen I, Collagen III, α-SMA, MMP 2 and MMP 9. Moreover, DSY repressed the phosphorylation of AKT, MAPKs and Smad3. In addition, DSY reduced inflammation and suppressed the mRNA expressions of IL-1β, IL-6, TNF-α, COX2 and iNOS in MF rats. ConclusionsOur study demonstrated that DSY prevented MF in vivo, the action of which was probably via reducing extracellular matrix organization, focal adhesion ECM-receptor interaction and inflammation by regulating TGF-β mediated PI3K/AKT, MAPK and Smad signaling pathways.

Pharmacokinetics of the material basis compounds of Tianshu capsule in treating migraine

Ethnopharmacological relevanceTianshu capsule (TSC) is a traditional Chinese medicine (TCM) preparation and has significant clinical effect on migraine. The composition characteristics of TSC formula are worth exploring for the treatment of migraine. Aim of the studyIdentify the compounds in vivo after oral administration of TSC, Gastrodin (GAS), ferulic acid, senkyunolide G and senkyunolide I in the blood of healthy and migraine rats were used as representative compounds for time-dependent processes investigation. And we focus on explaining the characteristics of TSC treatment on migraine from a pharmacokinetic perspective. Materials and methodsIn this study, Ultra-high performance liquid chromatography with Orbitrap tandem mass spectrometry (UPLC-Orbitrap-MS/MS) system were used to detect the compounds in rat plasma and brain after oral administration of TSC. A sensitive, selective and reliable ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-TQ-MS/MS) system method was established for the simultaneous quantitative analysis of multi-components and evaluate the pharmacokinetic behavior of four main compounds. ResultsA total of 46 compounds were significantly identified in vivo of rat, including 35 compounds in plasma, 7 compounds only in brain and 4 compounds both in plasma and brain. The quantitative method has been confirmed to be feasible for multi-components content determination, pharmacokinetic parameters indicated that the four compounds absorbed into blood rapidly, and senkyunolide I also cross the blood-brain barrier (BBB) quickly into brain. GAS has relatively high concentrations in plasma, and the parameters AUC0-t, CLz/F displayed significant differences between the normal and migraine rats. And the AUC0-t and Cmax of GAS and ferulic acid exhibited dose-dependent way. Conclusionswe added compounds for the pharmacokinetic study of TSC, providing powerful help for clinical medication in the treatment of migraine.

Erythrocyte Selenium as a Potential Key Indicator for Selenium Supplementation in Low-Selenium Populations: A Selenium Supplementation Study Based on Wistar Rats.

Selenium (Se) is an essential trace element for maintaining human health, with significant antioxidant and immunoregulatory functions. Inadequate Se intake may be associated with Keshan disease, Kashin-Beck disease, and hypothyroidism. However, effective indicators for scientifically guiding Se supplementation in Se-deficient populations are still lacking. This study aims to explore the dynamic distribution of Se across various nutritional biomarkers and major organs in rats through a Se supplementation experiment, as well as the pairwise correlations between them, in order to identify reliable nutritional indicators for evaluating Se levels in the body. Se levels in hair, blood, and major tissues and organs were determined by atomic fluorescence spectrometry, and glutathione peroxidase (GSH-Px) levels were measured using an ELISA. Se supplementation significantly increased Se levels in rat blood, hair, and major organs, as well as GSH-Px levels in blood. Se primarily accumulated in the liver and kidneys, followed by myocardium, spleen, and muscles. Serum and plasma Se were found to be the best indicators of short-term Se intake, while erythrocyte Se levels showed a stronger correlation with Se levels in tissues and organs, making it a better marker for assessing long-term Se nutritional status compared to hair Se. This study demonstrates the potential of erythrocyte Se levels as an indicator for evaluating long-term Se nutritional status, providing scientific evidence for Se nutritional assessments.

Effect of Rhei Radix Et Rhizome on treatment of polycystic ovary syndrome by regulating PI3K/AKT pathway and targeting EGFR/ALB in rats

Ethnopharmacological relevanceAbnormal endocrine metabolism caused by polycystic ovary syndrome (PCOS) poses a serious risk to reproductive health in females. According to Traditional Chinese Medicine (TCM) theories, the leading causes of PCOS include turbid phlegm, blood stasis and stagnation of liver Qi. Rhei Radix Et Rhizome is widely used in TCM to attack stagnation, clear damp heat, relieve fire. Rhei Radix Et Rhizome is an important part of the TCM formulas for the treatment of PCOS, which has a long history of medicinal use. However, the specific effect and mechanisms of Rhei Radix Et Rhizome on PCOS have yet to be elucidated. Aim of the studyThe object of this study aimed to investigate the effect and its pharmacological mechanism of Rhei Radix Et Rhizome on the treatment of polycystic ovary syndrome. MethodsPCOS was induced in female Sprague Dawley (SD) rats by administering letrozole (1 mg/kg, per orally, p.o.) for 21 days, then treated with Rhei Radix Et Rhizome at doses of 0.6 g/kg or 1.2 g/kg. Rats weight, blood glucose and estrus period are measured, and serum hormone include free testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and ovarian lesions were observed to determine the effects of Rhei Radix Et Rhizome. Network pharmacology and molecular docking predicted the targets of Rhei Radix Et Rhizome on PCOS. Epidermal growth factor receptor (EGFR), albumin (ALB), PI3K and P-AKT/AKT protein expression levels in ovarian tissues were assessed by Western blot. ResultsRhei Radix Et Rhizome reduce abnormal weight and fasting blood glucose induced by letrozole (n = 5, p < 0.01), and improve the disturbed estrus cycle, reduce T, LH levels and LH/FSH ratio of PCOS rats (n = 4, p < 0.01). In addition, it alleviates the polycystic changes of ovaries in PCOS rats and reduces ovarian histopathological damage (n = 4, p < 0.01). Additionally, the core active components of Rhei Radix Et Rhizome for PCOS include Sennoside D_qt, Procyanidin B-5,3′-O-gallate, and Mutatochrome, which strongly bind to core therapeutic targets ALB and EGFR. Furthermore, the treatment reduces the increase of EGFR and ALB induced by letrozole (n = 4, p < 0.01). KEGG pathway enrichment analysis highlights endocrine resistance and prolactin signaling pathway, in both of which the PI3K/AKT pathway plays a crucial role. Our results show Rhei Radix Et Rhizome rescue the abnormal expression of PI3K/AKT pathway in PCOS rats (n = 4, p < 0.01). However, no significant dose-dependent relationship was observed in the tested dose range for the above experiments. ConclusionThese findings suggest that Rhei Radix Et Rhizome can regulate the PI3K/AKT pathway and target EGFR and ALB to treat polycystic ovary syndrome in rats. This study provides a scientific basis for the use of Rhei Radix Et Rhizome in the treatment of PCOS and highlights its potential mechanism through modulation of the PI3K/AKT pathway.

High-performance electrochemical detection for trace minocycline based on boron-doped-diamond/carbon nanowalls biosensor

Highly sensitive electrochemical detection of trace minocycline (MCL), one of the most important antibiotics, presents a challenge due to the lack of suitable substrates and low detection efficiency. In this work, an electrochemical biosensor with an electrode consisting of boron-doped diamond (BDD) and carbon nanowalls (CNWs) (named BDD/CNWs) is designed for the detection of MCL. The electrochemical behavior of MCL at varying scan rates suggests an adsorption-controlled mechanism dominating the process on the BDD/CNWs electrode. The sensor demonstrates good linearity in the range from 0.02 to 100 μM and achieved a limit of detection as low as 9.8 nM, ascribed to the synergistic effect of the combination of BDD with CNWs, which provide a broad potential window (2.63 V), more reaction sites, and low charge transfer resistance (87 Ω) from the electrode. Examinations of trace MCL in citric acid-phosphate buffer, tap water, rat blood, as well as interference tests, reveal high sensitivity, specificity, stability, and repeatability of the BDD/CNWs electrode, which shows promise for practical applications.

Steroids' Neuroprotective Potential in Severe Cerebral Venous Thrombosis: Experimental and Clinical Exploration of NLRP3 Inflammasome Inhibition.

NLRP3 inflammasome-related inflammation might play an important role in the pathophysiology of severe CVT. The use of steroids as anti-inflammatory agents in improving severe CVT prognosis remains controversial. A total of 94 male Sprague-Dawley rats were used. We evaluated the dynamic and association between NLRP3 inflammasome in brain, blood, and CSF and severity in severe CVT rats and/or patients. We also explored the effect of steroids on NLRP3 activation, neurological injury, and CSF circulation disturbance after CVT in animals and/or patients. In rats, compared with the sham group, NLRP3-related factors rose on day 1, peaked on day 2 (NLRP3, Sham: 0.79 ± 0.22; day 2: 1.25 ± 0.08, p < 0.01; pro-Caspase-1, Sham: 0.58 ± 0.13, day 2: 1.20 ± 0.44, p < 0.05; GSDMD, Sham: 0.94 ± 0.22, day 2: 1.72 ± 0.46, p < 0.05; pro-IL-1β, Sham: 0.74 ± 0.15, day 2: 1.35 ± 0.09, p < 0.01), decreased on day 7 in rats (n = 4 per group). Thrombus (Sham: 0.00 ± 0.00, day 2: 3.44 ± 0.70, p < 0.0001), infarct size (Sham: 0.00 ± 0.00, day 2: 11.99 ± 6.26, p < 0.01) and neurological deficits appeared similar trend. In 50 patients, serum NLRP3 and IL-6 levels correlated positively with NIHSS (r = 0.4273, p = 0.0020; r = 0.4938, p = 0.0029) and mRS (r = 0.5349, p = 0.0125; r = 0.6213, p = 0.026), while CSF IL-6 correlated positively with mRS on admission (r = 0.5349, p = 0.0125). Compared with baseline, NLRP3 (0.36 (0.36, 0.36) vs. 0.41 (0.37, 0.84), p < 0.0001) and IL-6 decreased (4.06 ± 1.48 vs. 12.03 ± 7.80, p < 0.05), accompanying by improvement of neurological deficits and CSF circulation (all p < 0.01) after steroids therapy in severe CVT patients at discharge and 3 months follow-up. No significant steroid-related adverse effects were observed. Short-term steroid therapy may improve prognosis of severe CVT by suppressing NLRP3 inflammasome-related inflammation.

Active immunization against gonadotropin-releasing hormone enhances the generation of B cells but does not affect their colonization in peripheral immune organs in male rats

Active immunization against gonadotropin-releasing hormone (GnRH) affects the immune system by inhibiting testosterone production. Our previous study investigated the effects of GnRH immunization on thymic T-cell generation, migration, and colonization in peripheral immune organs. However, the mechanisms by which GnRH immunization influences B cell generation and the characteristics of B cell colonization in peripheral immune organs remain unclear. Herein, GnRH immunization enhanced B cell generation by reducing apoptosis. GnRH immunization did not markedly affect the cell cycle of bone marrow cells, B cell development-related signaling molecules, or the percentage of B cells in the blood, spleen, or inguinal lymph nodes. After testosterone supplementation in GnRH-immunocastrated rats, the generation of B cells in the bone marrow was significantly reduced, and the apoptosis of B cells was remarkably increased. Testosterone did not significantly affect the cell cycle of bone marrow cells or the proportion of B cells in the blood, spleen, or inguinal lymph nodes of the GnRH-immunocastrated rats. Overall, these results clarify the mechanisms related to B cell expansion in the bone marrow and the settlement characteristics of B cells in peripheral immune organs after GnRH immunization.

Histomorphometric changes in pituitary gonadotropic endocrinocytes when exposed to dark deprivation

Aim. To assess the effect of 30-day dark deprivation on functional and histomorphometric changes in adenohypophysis gonadotropic endocrinocytes and their reversibility in mature male rats.Materials and methods. Mongrel white male rats (n = 36) weighing 365–375 g at 4 months of age were randomly divided into three groups (each n = 12). For 30 days the control group was in automatic light-dark mode 12/12, and the rats of experimental groups 1 and 2 were in round-the-clock artificial lighting (24/0, 300 Lux), then the rats of group 2 were returned to 12/12 mode for the next 14 days. In the animals of the control and group 1 during their lifetime on the 31st day, and in group 2 on the 45th day, blood was taken from the abdominal aorta and levels of follicle-stimulating (FSH) and luteinizing (LH) hormones, melatonin, and Klotho protein were determined an enzyme-linked immunosorbent assay and immunoassay and after which they were removed from the experiment by decapitation. Postmortem histological and immunohistochemical examination of the pituitary gland was done using rabbit polyclonal antibodies targeting caspase-3 and Klotho protein, as well as morphometry. Statistical data processing was performed using the Kruskal-Wallis test with post-hoc Dunn’s test.Results. Light desynchronization in the form of 30 days of dark deprivation increased FSH and LH levels and decreased melatonin and Klotho protein levels in the blood of male rats; increased gonadotropic endocrine cell area, volume, and perimeter by 23.1% (p &lt; 0.001), 48.7% (p &lt; 0.001), and 10.9% (p &lt; 0.001), respectively; and increased nucleus area, volume, and perimeter by 16%, 11.7%, and 2.5%, respectively. An immunohistochemical study showed an increase in the specific area of caspase-3-immunoreactive gonadotropic endocrinocytes by 25.2% without obvious morphological signs of apoptosis, and a decrease in the expression of Klotho protein by 25.7%. All indicators were reversible, the levels of FSH and Klotho protein in the blood of animals almost reached their initial values after 14 days of restoration of the light-dark cycle 12/12.Conclusion. Dark deprivation for 30 days in male rats induced reversible processes of accelerated aging and apoptosis in cells, as evidenced by changes in the expression of aging markers in gonadotropic endocrinocytes and levels of gonadotropic hormones in the blood. When the light-dark mode is restored, the levels of FSH and Klotho protein normalize as early as 14 days.