Days Of Continuous Administration Research Articles

Effect of CYP3A5*3 genotype on exposure and efficacy of quetiapine: A retrospective, cohort study

BackgroundThe involvement of cytochrome P450 3A5 (CYP3A5) in the metabolism of quetiapine has been proposed, though conclusive evidence is lacking. This study aimed to quantitatively assess the impact of CYP3A5 genetic variability on quetiapine exposure in a Chinese patient population. MethodsPatient data were retrospectively collected from the database of the Mental Health Centre at the First Hospital of Hebei Medical University, covering the period from September 1, 2019, to July 1, 2023. The study included patients genotyped for CYP3A5 who were treated with quetiapine. Inclusion criteria for the analysis of pharmacokinetic parameters, such as serum concentrations of the drug and its metabolites, included oral administration of quetiapine, availability of information on the prescribed daily dose and concomitant medications, and the determination of steady-state blood levels at the time of sampling (after at least 3 days of continuous administration at the same dose). Exclusion criteria comprised polypharmacy with known CYP3A4 inducers or inhibitors, as well as patients with hepatic or renal insufficiency. The primary endpoint was the exposure to quetiapine and N-dealkylquetiapine, measured using dose-corrected concentrations (C/D). The secondary endpoint was the metabolism of quetiapine to N-dealkylquetiapine, assessed by the ratio of metabolite to parent drug concentrations. The third endpoint is the differences in adverse reactions, QTc intervals, and biochemical parameters among patients with different CYP3A5 genotypes. ResultBased on the inclusion and exclusion criteria, clinical data from 207 patients were ultimately included in the study. Of these, 20 patients had the CYP3A5*1/*1 genotype, 78 had the CYP3A5*1/*3 genotype, and 109 had the CYP3A5*3/*3 genotype.The CYP3A5*3 variant was found to significantly impact the metabolism of quetiapine. The C/D values for both quetiapine and dealkylated quetiapine were notably higher in individuals with the *3/*3 genotype compared to those with the *1/*1 and *1/*3 genotypes (P1 <0.001 and P2 = 0.002, respectively). A comparison of the variability in metabolic ratios among different genotype groups revealed no significant difference (P = 0.067). However, a post hoc analysis indicated that the metabolic ratio in poor metabolizers was significantly lower than that in intermediate metabolizers (P = 0.021). The analysis of adverse reaction incidence and QTc intervals among different genotypes showed no statistically significant differences (P = 0.652, P = 0.486). However, comparison of biochemical parameters across different genotype groups revealed that alanine aminotransferase, uric acid, hemoglobin, and gamma-glutamyl transferase levels were significantly higher in patients with the CYP3A5*3/*3 genotype compared to those with the CYP3A5*1/*1 and CYP3A5*1/*3 genotypes. ConclusionThe results indicated that the genetic polymorphism of CYP3A5*3 significantly influences the metabolism of quetiapine. Specifically, carriers of the CYP3A5*3/*3 genotype exhibited higher blood levels of quetiapine, with a greater likelihood of these levels exceeding the therapeutic range. This finding underscores the need for clinicians to carefully monitor and potentially adjust the dosage for patients with this genotype to avoid adverse effects. This finding underscores the need for clinicians to pay special attention to the efficacy and occurrence of adverse reactions when prescribing quetiapine to patients carrying the CYP3A5*3/*3 genotype.

Ultrafiltration of Angelicae Sinensis Radix and Astragali Radix inhibits ferroptosis and improves pulmonary fibrosis in rats by regulating Nrf2/xCT/GPX4 signaling pathway

This study aims to investigate the mechanism of ferroptosis mediated by the nuclear factor-E2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11, also known as xCT)/glutathione peroxidase 4(GPX4) signaling pathway in radiationinduced pulmonary fibrosis and the intervention effect of Angelicae Sinensis Radix(ASR) and Astragali Radix(AR) ultrafiltration extract. Fifty Wistar rats were randomly divided into five groups, with 10 rats in each group. Except for the blank group without radiation, the rats in each group were anesthetized and subjected to a single local chest irradiation of 40 Gy X-rays once to establish a rat model of radiation-induced pulmonary fibrosis. After radiation, the rats in the intervention groups were orally administered with ASR-AR ultrafiltration extract at doses of 0. 12, 0. 24, and 0. 48 g·kg~(-1), respectively, once a day for 30 days. After 30 days of continuous administration, the levels of oxidative stress indicators superoxide dismutase(SOD) activity, reduced glutathione(GSH),malondialdehyde(MDA), and ferrous ion(Fe~(2+)) in lung tissues of each group were detected by colorimetry. Immunofluorescence was used to detect reactive oxygen species(ROS) fluorescence expression in lung tissues. Hematoxylin-eosin(HE) and Masson staining were performed to observe pathological changes in lung tissues. Immunohistochemistry and Western blot were used to detect the expression levels of Nrf2/xCT/GPX4 signaling pathway and fibrotic proteins in lung tissues. The results showed that compared with the results in the blank group, the levels of Fe~(2+) and MDA in the model group increased, while SOD activity and GSH levels decreased,and ROS levels increased. HE and Masson staining results showed that the structure of lung tissue was seriously damaged, the pulmonary interstitium was significantly proliferated, the alveoli collapsed and consolidated severely, and there were more inflammatory cell aggregates and collagen fiber deposits. Transmission electron microscopy showed that the degree of lung tissue damage in the model group was relatively high, with increased, smaller, and disorganized damaged mitochondria, irregular morphology, shallow matrix,most mitochondria ruptured and shortened, mildly expanded, some mitochondria with increased electron density of the matrix, partial mitochondrial outer membrane rupture, and characteristic changes of ferroptosis-specific mitochondria. Immunohistochemistry showed that the expression of transferrin receptor protein 1(TFR1) in lung tissues was significantly increased, while the expression of GPX4,ferritin heavy chain 1(FTH1), Nrf2, and xCT was significantly decreased. Western blot showed that the expression of α-smooth muscle actin(α-SMA) and collagen Ⅰ protein increased. Compared with the model group, the intervention group with ASR-AR ultrafiltration extract significantly improved lipid peroxidation and antioxidant-related indicators, decreased Fe~(2+) levels, alleviated fibrosis, and decreased the expression of TFR1, α-SMA, and collagen Ⅰ proteins in lung tissues, while increased the expression of GPX4, FTH1, Nrf2, and xCT proteins. In summary, ASR-AR ultrafiltration extract has an ameliorative effect on radiation-induced pulmonary fibrosis, and its mechanism may involve the inhibition of ferroptosis by regulating the Nrf2/xCT/GPX4 signaling pathway.

Amlodipine Protects against Methotrexate-Induced Acute Kidney Injury in Rats

Background Methotrexate (MTX) is a commonly used chemotherapy drug with known nephrotoxic effects, including the potential for acute kidney injury. However, the precise mechanism through which MTX induces nephrotoxicity remains unclear, though oxidative stress and direct toxic effects on renal tubules are believed to play key roles. Recent studies suggest that calcium channel blockers may offer promise in slowing down the progression of chronic kidney diseases. Objective The purpose of this study was to explore the potential of Amlodipine, a calcium channel blocker, to alleviate acute kidney injury caused by the administration of MTX in rats. Methods Three groups of twenty-four male Wistar rats were randomly assigned: Group 1—the control group was given normal saline orally. Group II, underwent five days of continuous administration of a single intraperitoneal (IP) dosage of 20 mg/kg MTX. The same dosage of MTX was given to Group III followed by an oral dose of Amlodipine at 5 mg/kg over the same period. Upon completion of the experiment, serum biochemical parameters, renal damage markers, oxidative stress, inflammatory markers, and kidney tissue histology were assessed. Results The results indicate that MTX administration significantly increased the levels of serum biochemical parameters, renal damage markers, inflammatory markers, oxidative stress markers, and induced alterations in kidney histology. However, the administration of Amlodipine following MTX treatment protected against these changes. Conclusion Amlodipine exhibits therapeutic potential in mitigating MTX-induced kidney injury in rats and its associated side effects.

Formononetin reverses Treg/Th17 imbalance in immune-mediated bone marrow failure mice by regulating the PI3K/Akt signaling pathway

BackgroundSevere aplastic anemia (SAA) is a syndrome of bone marrow failure which is life-threatening. Recent studies have demonstrated that CD4 + T cell subsets, including T regulatory (Treg) and T helper 17 (Th17) cells, play a pivotal role in the pathogenesis of SAA. Formononetin (FMN) is a natural compound extracted from the traditional Chinese medicine Huangqi, which has the ability to regulate the imbalance of Treg/Th17 cells in some inflammatory diseases. Nevertheless, the therapeutic effect of FMN in SAA has yet to be definitively established. Therefore, the objective of this research was to investigate the effect of FMN on SAA and elucidate its underlying mechanism.MethodsIn vivo experiments, the mice were divided into the following five groups: control, model, low-dose FMN, high-dose FMN, and positive control cyclosporine A group. The immune-mediated bone marrow failure (BMF) mouse model was established by the total body X-ray radiation and lymphocyte infusion. After 10 days of continuous administration of FMN, the numbers of Treg/Th17 cells in the bone marrow and spleen were assessed by flow cytometry. The protein expressions of PI3K/Akt pathway in the bone marrow and spleen was assessed by immunohistochemistry and western blotting. In vitro, the impact of FMN on the differentiation of naive CD4 + T cells into Treg cells was investigated by flow cytometry and ELISA.ResultsIn comparison with the control group, the model group showed a reduction in bone marrow nucleated cells, a significant decrease in peripheral blood cells, and an altered CD8 + /CD4 + T cell ratio. These findings indicate the successful establishment of a mouse model of immune-mediated BMF. After FMN treatment, there were the increased levels of red blood cells and hemoglobin. In addition, FMN mitigated the bone marrow destruction and restored the CD8 + /CD4 + T cell ratio. Furthermore, in comparison with the control group, the model group showed the decreased levels of Treg cells and the increased levels of Th17 cells. After FMN treatment, there was a significantly increased number of Treg cells and a decreased number of Th17 cells. Additionally, FMN remarkably down-regulated the expression levels of PI3K and Akt proteins in immune-mediated BMF mice.ConclusionsFMN alleviates immune-mediated BMF by modulating the balance of Treg/Th17 cells through the PI3K/Akt signaling pathway.

The expression of ProBDNF and its high affinity receptor P75NTR in the neurons of emotion-related brain regions of post-stroke depression rats

ObjectiveTo investigate the expression of the precursor of brain-derived neurotrophic factor (proBDNF) and its high-affinity receptor p75NTR in neurons of emotion-related brain areas (prefrontal cortex, hippocampus, and amygdala) in rats with post-stroke depression (PSD), and to explore the expression levels of proBDNF and p75NTR in neurons of emotion-related brain areas by injecting tissue plasminogen activator (t-PA) into the lateral ventricle of PSD rats, this significantly improved the stress-induced depression-like behavior,thus further validating the above results. MethodsRats were randomly divided into four groups: a normal control group (n = 8), a depression group (n = 8), a stroke group (n = 8), and a PSD group (n = 8). The rat model of stroke was established by thread embolism, and the PSD animal model was induced by chronic unpredictable mild stress (CUMS) and solitary feeding. Behavioral tests were conducted, including weight measurement, open field tests, and sucrose preference tests. Immunofluorescence double labeling was used to detect the expression of proBDNF and p75NTR in neurons of emotion-related brain regions in the PSD rat model. Four weeks after CUMS treatment, the PSD group was selected. Rats were infused with t-PA (3 μg dissolved in 6 μL saline, Boehringer Ingelheim), proBDNF (3 μg dissolved in 6 μL saline, Abcam), or equal-volume NS once per day for 7 consecutive days using the syringe pump connecting to injection needles. After 7 days of continuous administration, animal behavior was assessed through scoring, and the expression of proBDNF and p75NTR in the emotion-related brain regions of the PSD rat model was detected using immunofluorescence double labeling. ResultsCompared with the normal control group and the stroke group, the body weight, sucrose water consumption, and vertical movement distance in the PSD group were significantly lower (P < 0.05). In contrast, when compared with the proBDNF injection group and saline injection group, the weight, sucrose water consumption, field horizontal movement, and vertical movement distance of the t-PA injection group significantly increased after PSD lateral ventricle intubation.Double immunofluorescence revealed a higher neuronal expression of proBDNF as well as p75NTR in the prefrontal cortex and hippocampus of PSD rats compared to control animals (P < 0.05). In the amygdala, the expression levels of proBDNF and P75NTR were significantly reduced in the PSD group compared to the control group (P < 0.05). The results of the expression levels of proBDNF and P75NTR in the emotion-related brain regions of PSD rats injected with t-PA showed that proBDNF and P75NTR was significantly reduced in the prefrontal cortex, hippocampus, and amygdala of PSD rats compared to those of the NS and proBDNF groups (P < 0.05). ConclusionsThe increased expression of the brain-derived neurotrophic factor precursor proBDNF and its receptor p75NTR in neurons of emotion-related brain regions may play an important role in the pathogenesis of PSD.t-PA reduced the expression of proBDNF and its receptor p75NTR in neurons emotion-related brain regions and significantly improved the stress-induced depression-like behavior. Therefore, it is reasonable to assume that exogenous injection of t-PA may alleviate the depressive symptoms of PSD patients.Reducing the expression of proBDNF by injecting t-PA may provide a novel therapeutic approach for the treatment of stress-related mood disorders.

Molecular Mechanisms of Phyllanthus urinaria in Cerebral Ischemia-Reperfusion Injury

Phyllanthus urinaria (P. urinaria), a prominent herb in traditional Zhuang medicine, is characterized by its heat clearance, detoxification, diuresis, and detumescence. This study investigates the therapeutic effects and molecular mechanisms of P. urinaria on cerebral ischemia-reperfusion injury (CIRI). 60 adult SD rats were randomly divided into four groups: a sham operation group, a model group of right middle cerebral artery occlusion (MCAO), a low-dose group treated with P. urinaria at 5 g/kg, and a high-dose group treated with P. urinaria at 10 g/kg, each consisting of 15 rats. After seven days of continuous administration, a rat model of the right MCAO was established to simulate CIRI. Nerve function was assessed using the Longa scoring method, the cerebral infarction area was evaluated with TTC staining, and cytokine levels (IL-1β, TNF-α, IL-6) were measured via ELISA. PI3K and AKT protein expression in brain tissues was analyzed by Western blot. The results revealed a significant improvement in neurological function scores in both low and high-dose P. urinaria groups compared to the MCAO model group. Treatment with P. urinaria led to a notable reduction in cerebral infarction area after 72 hours of cerebral ischemia. The levels of inflammatory cytokines (IL-1β, TNF-α, IL-6) were decreased in both treatment groups compared to the MCAO model group. There was a significant increase in the expression of PI3K and AKT proteins in both treatment groups. In conclusion, P. urinaria exhibits a protective effect against CIRI by inhibiting inflammatory responses, improving neurological function, and reducing brain injury.

Distribution and Elimination of Deltamethrin Toxicity in Laying Hens.

Deltamethrin, an important pyrethroid insecticide, is frequently detected in human samples. This study aims to assess the potential effects of deltamethrin on human health and investigate the patterns of residue enrichment and elimination in 112 healthy laying hens. These hens were administered 20 mg·kg-1 deltamethrin based on their body weight. Gas chromatography-mass spectrometry (GC-MS) was used to investigate the residue enrichment pattern and elimination pattern of deltamethrin in the hens. The results indicated a significant increase in the concentration of deltamethrin in chicken manure during the treatment period. By the 14th day of administration, the concentration of deltamethrin in the stool reached 13,510.9 ± 172.24 μg·kg-1, with a fecal excretion rate of 67.56%. The pulmonary deltamethrin concentration was the second highest at 3844.98 ± 297.14 μg·kg-1. These findings suggest that chicken feces contain substantial amounts of deltamethrin after 14 days of continuous administration, and that it can easily transfer to the lungs. After 21 days of drug withdrawal, the residual concentration of deltamethrin in the fat of laying hens was 904.25 ± 295.32 μg·kg-1, with a half-life of 17 days and a slow elimination rate. In contrast, the lungs showed relatively low elimination half-lives of 0.2083 days, indicating faster elimination of deltamethrin in this tissue. These results highlight differences in the rate of deltamethrin elimination in different tissues during drug withdrawal. The fat of laying hens exhibited the highest residue of deltamethrin and the slowest elimination rate, while the lungs showed the fastest elimination rate. Moreover, deltamethrin was found to accumulate in the edible tissues of eggs and laying hens, suggesting that humans may be exposed to deltamethrin through food.

Exploring the Mechanism of Chang-An-Kang on Active Ulcerative Colitis Rats Based on NF-κB/STAT3 Signaling Pathway

Objectives The objective of this study was to investigate the therapeutic effect and mechanism of Chang-An-Kang on ulcerative colitis rats. Materials and Methods The rat model of ulcerative colitis induced by 6% glacial acetic acid was established. The corresponding therapeutic drugs were given 24 h after the model was established. The pathological morphology of colon tissue was observed after 21 days of continuous administration. The levels of interleukin-6 (IL-6), interleukin-7 (IL-7), interleukin-10 (IL-10), and tumor necrosis factor-alpha (TNF-α) were measured by ELISA kit. The expression levels of NF-κB and STAT3 proteins in colon tissue were detected by IHC. Results The results showed that the changes in the pathological and biochemical indexes of the rat colon tissue indicate that the model was successfully established. Chang-An-Kang could significantly reduce rat colon tissue damage, decrease the content of IL-6, IL-7, and TNF-α, and increase the content of IL-10. The NF-κB and STAT3 protein expression levels were decreased. Conclusion By increasing the content of anti-inflammatory factor IL-10, decreasing the content of pro-inflammatory factors IL-6 and TNF-α, and inhibiting the expression of NF-κB and STAT3 proteins, Chang-An-Kang can reduce inflammatory infiltration. It has an obvious therapeutic effect on acetic acid ulcerative colitis in rats.

Intermittent Sargramostim Administration Expands Proliferating Naïve T Cells, Tregs, HLA-DR+ PD-L1+ Monocytes and Myeloid-Derived Suppressor Cells: Results from a Randomized Placebo-Controlled Clinical Trial of GM-CSF in Patients with Peripheral Artery Disease

Introduction: Sargramostim (rhuGM-CSF; Leukine ®) is an FDA-approved drug known for mobilizing hematopoietic progenitor cells and expediting hematopoietic reconstitution in patients with acute myeloid leukemia (AML) as well as after autologous and allogeneic bone marrow transplantation. While GM-CSF has been explored as an adjuvant for cancer vaccines due to its ability to promote dendritic cell differentiation and maturation, continuous daily administration of sargramostim has been associated with leukocytosis and adverse side effects. To validate and expand upon our previously published findings that thrice-weekly sargramostim administration in patients with severe peripheral artery disease (PAD) is both well-tolerated and beneficial for claudication symptoms, we conducted a placebo-controlled randomized clinical trial to assess the efficacy of an extended dosing regimen of sargramostim in PAD patients. Data regarding progenitor mobilization kinetics and claudication symptoms will be presented elsewhere. In this analysis, we investigated the impact of intermittent sargramostim dosing on immune cell subsets in the bloodstream, aiming to support the use of this dosing schedule for enhancing adaptive immunity. Methods: Symptomatic atherosclerotic PAD patients were enrolled and randomly assigned to receive either 500 μg/day of sargramostim or placebo via self-administered subcutaneous injections thrice weekly for 3 weeks, for two separate courses 3 months apart. The study was IRB-approved with adverse events and safety signals monitored by an independent Data Safety Monitoring Board. A total of 61 patients (40 receiving sargramostim and 21 receiving placebo) participated in the immune-monitoring sub-study. Blood mononuclear cells were collected at various time points and cryopreserved. Thawed peripheral blood mononuclear cells (PBMCs) were stained using immune cell-specific antibody cocktails, and analyzed by flow cytometry. Statistical significance was determined using the Student's t-test. Results: No dose-limiting leukocytosis or drug-related side effects were observed. As expected, the numbers of white blood cells (WBCs) increased by an average of 68% in the blood of sargramostim-treated patients compared to placebo-treated patients, with peak values reached after 3 weeks of intermittent study drug administration, followed by return to baseline before the next treatment course. Absolute counts of CD3+, CD4+, and CD8+ T cells, B cells, and CD14+ monocytes significantly increased one and two weeks after starting intermittent sargramostim treatment compared to the control group. Notably, sargramostim-treated patients exhibited higher levels of proliferating naïve, Ki-67+ CD4+ T cells (106.83/uL ± 70.28) compared to placebo-treated patients (44.94/uL ± 33.84, p=0.001), as well as increased numbers of naïve Ki-67+ CD8+ T cells (33.05/uL ± 25.43 after sargramostim versus 15.77/uL ± 15.73 for placebo, p=0.01) (Fig. 1, A). However, sargramostim treatment did not influence the quantities of central memory CD4+ and CD8+ T cells. Besides the heightened proliferation of naïve T cells, sargramostim treatment also resulted in a significant increase in the numbers of CD3+CD4+CD25+FoxP3+ Tregs (Fig. 1, A), as well as PD-1-expressing CD4+ and CD8+ T cells, compared to the control group. Moreover, sargramostim treatment led to elevated levels of CD14+HLA-DR+ and CD14+PDL-1+ monocytes, as well as myeloid-derived suppressor cells, 2 and 3 weeks after initiation of treatment, when compared to the control group (Fig. 1, B). Conversely, sargramostim treatment had no impact on the levels of gamma-delta T cells, natural killer (NK) cells, plasmacytoid dendritic cells, or classical dendritic cells. Conclusions: Our findings demonstrate that intermittent dosing with sargramostim is well-tolerated and induces significant but transient increases in the numbers and activation status of T cells in the bloodstream. Sargramostim treatment also increased numbers of proliferating naïve T cells, Tregs, and certain subsets of myelo-monocytic cells with immunosuppressive phenotypes. These results support the use of intermittent sargramostim dosing to avoid excessive leukocytosis and associated adverse effects. Further investigation is warranted to elucidate the clinical implications of these findings in the context of immune modulation.

Protective effect of Liujing Toutong Tablets on rats with permanent cerebral ischemia via NF-κB signaling pathway

This study investigated the neuroprotective effects and underlying mechanism of Liujing Toutong Tablets(LJTT) on a rat model of permanent middle cerebral artery occlusion(pMCAO). The pMCAO model was established using the suture method. Eighty-four male SPF-grade SD rats were randomly divided into a sham operation group, a model group, a nimodipine group(0.020 g·kg~(-1)), and high-, medium-, and low-dose LJTT groups(2.8, 1.4, and 0.7 g·kg~(-1)). The Longa score, adhesive removal test and laser speckle contrast imaging technique were used to evaluate the degree of neurological functional impairment and changes in local cerebral blood flow. The survival and mortality of rats in each group were recorded daily. After seven days of continuous administration following the model induction, the rats in each group were euthanized, and brain tissue and blood samples were collected for corresponding parameter measurements. Nissl staining was used to examine pathological changes in brain tissue neurons. The levels of tumor necrosis factor-alpha(TNF-α), interleukin-6(IL-6), IL-1β, vascular endothelial growth factor(VEGF), calcitonin gene-related peptide(CGRP), beta-endorphin(β-EP), and endogenous nitric oxide(NO) in rat serum were measured using specific assay kits. The entropy weight method was used to analyze the weights of various indicators. The protein expression levels of nuclear factor kappa-B(NF-κB), inhibitor kappaB alpha(IκBα), phosphorylated IκBα(p-IκBα), and phosphorylated inhibitor of NF-κB kinase alpha(p-IKKα) in brain tissue were determined using Western blot. Immunohistochemistry was used to detect the protein expression of chemokine-like factor 1(CKLF1) and C-C chemokine receptor 5(CCR5) in rat brain tissue. Compared with the sham operation group, the model group showed significantly higher neurological functional impairment scores, prolonged adhesive removal time, decreased cerebral blood flow, increased neuronal damage, reduced survival rate, significantly increased levels of TNF-α, IL-1β, IL-6, CGRP, and NO in serum, significantly decreased levels of VEGF and β-EP, significantly increased expression levels of NF-κB p65, p-IκBα/IκBα, and p-IKKα in rat brain tissue, and significantly upregulated protein expression of CKLF1 and CCR5. Compared with the model group, the high-dose LJTT group significantly improved the neurological functional score of pMCAO rats after oral administration for 7 days. LJTT at all doses significantly reduced adhesive removal time and restored cerebral blood flow. The high-and medium-dose LJTT groups significantly improved neuronal damage. The LJTT groups at all doses showed reduced levels of TNF-α, IL-1β, IL-6, CGRP, and NO in rat serum, increased VEGF and β-EP levels, and significantly decreased expression levels of NF-κB p65, p-IκBα/IκBα, p-IKKα, and CCR5 protein in rat brain tissue. The entropy weight analysis revealed that CGRP and β-EP were significantly affected during the model induction, and LJTT exhibited a strong effect in reducing the release of inflammatory factors such as TNF-α and IL-1β. LJTT may exert a neuroprotective effect on rats with permanent cerebral ischemia by reducing neuroinflammatory damage, and its mechanism may be related to the inhibition of the NF-κB signaling pathway and the regulation of the CKLF1/CCR5 axis. Additionally, LJTT may exert certain analgesic effects by reducing CGRP and NO levels and increasing β-EP levels.

Alleviation of ultraviolet-B radiation-induced photoaging using Saussurea medusa Maxim polysaccharide.

Saussurea medusa polysaccharide, the polysaccharide extract of Saussurea medusa Maxim, a traditional Chinese herbal medicine, is used to combat intense ultraviolet radiation, cold, and hypoxia in patients, as well as during drought. This polysaccharide has rich medicinal and ecological values. We aimed to determine whether saussurea medusa polysaccharides can reduce ultraviolet B (UVB)-induced skin photoaging. Seventy-five male Kunming mice were divided into five groups: control, UVB-only, UVB plus vitamin E (VE group), UVB plus saussurea medusa (2 g/kg), and UVB plus saussurea medusa (6 g/kg). The control group was irradiated with normal light, while the other four groups were subcutaneously administered 10 mL/kg/day D-galactose and irradiated with narrow-spectrum UVB for 40 min daily. From day 11, the VE group was administered 0.25 g/kg/day vitamin E, while the saussurea medusa intervention groups were administered 2 and 6 g/kg/day saussurea medusa polysaccharide. After 30 days of continuous administration, treatment with saussurea medusa polysaccharides was found to reduce UVB-induced skin photoaging in mice by elevating the levels of superoxide dismutase, glutathione peroxidase, and hydroxyproline (HYP), while reducing the level of MDA, and inhibiting the EGFR/MEK/ERK/c-Fos pathway. Overall, our findings suggest that treatment with saussurea medusa polysaccharides positively influences skin photoaging.

Biocompatibility and Safety Assessment of Combined Topical Ozone and Antibiotics for Treatment of Infected Wounds.

Wound infections with antibiotic-resistant bacteria, particularly the Gram-negative strains, pose a substantial health risk for patients with limited treatment options. Recently topical administration of gaseous ozone and its combination with antibiotics through portable systems has been demonstrated to be a promising approach to eradicate commonly found Gram-negative strains of bacteria in wound infections. However, despite the significant impact of ozone in treating the growing number of antibiotic-resistant infections, uncontrolled and high concentrations of ozone can cause damage to the surrounding tissue. Hence, before such treatments could advance into clinical usage, it is paramount to identify appropriate levels of topical ozone that are effective in treating bacterial infections and safe for use in topical administration. To address this concern, we have conducted a series of in vivo studies to evaluate the efficacy and safety of a portable and wearable adjunct ozone and antibiotic wound therapy system. The concurrent ozone and antibiotics are applied through a wound interfaced gas permeable dressing coated with water-soluble nanofibers containing vancomycin and linezolid (traditionally used to treat Gram-positive infections) and connected to a portable ozone delivery system. The bactericidal properties of the combination therapy were evaluated on an ex vivo wound model infected with Pseudomonas aeruginosa, a common Gram-negative strain of bacteria found in many skin infections with high resistance to a wide range of currently available antibiotics. The results indicated that the optimized combination delivery of ozone (4 mg h-1) and topical antibiotic (200 μg cm-2) provided complete bacteria eradication after 6 h of treatment while having minimum cytotoxicity to human fibroblast cells. Furthermore, in vivo local and systemic toxicity studies (e.g., skin monitoring, skin histopathology, and blood analysis) on pig models showed no signs of adverse effects of ozone and antibiotic combination therapy even after 5 days of continuous administration. The confirmed efficacy and biosafety profile of the adjunct ozone and antibiotic therapy places it as a strong candidate for treating wound infection with antimicrobial-resistant bacteria and further pursuing human clinical trials.

Total Glucosides of Paeony Regulate Immune Imbalance Mediated by Dermal Mesenchymal Stem Cells in Psoriasis Mice.

To investigate the therapeutic effects of total glucosides of paeony (TGP) on psoriasis based on the immunomodulatory effect of dermal mesenchymal stem cells (DMSCs). A total of 30 male BALB/c mice were divided into 6 groups (n=5 in each) by a random number table method, including control, psoriasis model (model, 5% imiquimod cream 42 mg/d), low-, medium- and high-dose TGP (50, 100, and 200 mg/kg, L, M-, and H-TGP, respectively), and positive control group (2.5 mg/kg acitretin). After 14 days of continuous administration, the skin's histopathological changes, apoptosis, secretion of inflammatory cytokines, and proportion of regulatory T cells (Treg) and T helper cell 17 (Th17) were evaluated using hematoxylin-eosin (HE) staining, TdT-mediated dUTP nick end labeling staining, enzyme-linked immunosorbent assay, and flow cytometry, respectively. DMSCs were further isolated from the skin tissues of normal and psoriatic mice, and the cell morphology, phenotype, and cycle were observed. Furthermore, TGP was used to treat psoriatic DMSCs to analyze the effects on the DMSCs immune regulation. TGP alleviated skin pathological injury, reduced epidermis layer thickness, inhibited apoptosis, and regulated the secretion of inflammatory cytokines and the proportion of Treg and Th17 in the skin tissues of psoriatic mice (P<0.05 or P<0.01). There was no significant difference in cell morphology and phenotype between control and psoriatic DMSCs (P>0.05), however, more psoriatic DMSCs remained in G0/G1 phase compared with the normal DMSCs (P<0.01). TGP treatment of psoriatic DMSCs significantly increased cell viability, decreased apoptosis, relieved inflammatory response, and inhibited the expression of toll-like receptor 4 and P65 (P<0.05 or P<0.01). TGP may exert a good therapeutic effect on psoriasis by regulating the immune imbalance of DMSCs.

Cerebroprotein hydrolysate attenuates neurodegenerative changes in Alzheimer's mice model via ferroptosis pathway.

Introduction: Cerebroprotein hydrolysate has been proven to improve cognitive function in patients with Alzheimer's disease (AD). We explored the safety and effectiveness of the clinical administration of oral cerebroprotein hydrolysate in AD, and possible mechanisms related to the neuronal ferroptosis pathway. Methods: Three-month-old male APP/PS1 double-transgenic mice were randomly divided into AD model (n = 8) and intervention (n = 8) groups. Eight non-transgenic wild-type (WT) C57 mice were used as age-matched controls. The experiments were started at the age of 6 months. The intervention group was then administered cerebroprotein hydrolysate nutrient solution (11.9mg/kg/day) via chronic gavage, the other groups received an identical volume of distilled water. Behavioural experiments were performed after 90days of continuous administration. Serum and hippocampal tissues were then collected for histomorphological observation, tau and p-tau expression, and ferroptosis markers analysis. Results: Cerebroprotein hydrolysate simplified movement trajectories and shortened escape latencies of APP/PS1 mice in the Morris water maze test. Neuronal morphologies were restored in hippocampal tissues on haematoxylin-eosin staining. In the AD-model group, Aβ protein and p-tau/tau expression levels were elevated, plasma Fe2+ and malondialdehyde levels were elevated, GXP4 protein expression and plasma glutathione levels declined than controls. All indices improved after cerebroprotein hydrolysate intervention. Conclusion: Cerebroprotein hydrolysate improves learning and memory function, alleviates neuronal damage, and reduces the deposition of pathological AD markers in AD mice, which may be related to the inhibition of neuronal ferroptosis.

Safety, efficacy and pharmacokinetics of BPI-9016M in c-MET overexpression or MET exon 14 skipping mutation patients with locally advanced or metastatic non-small-cell lung cancer: a phase Ib study

BackgroundAs a potential target receptor tyrosine kinase, mesenchymal-epithelial transition factor (MET) exhibits high aberrant expression across various tumors. This study aimed to evaluated the safety, tolerability, efficacy and pharmacokinetics (PK) of BPI-9016M, a novel tyrosine kinase inhibitor (TKI) targeting c-MET, in c-MET overexpression or MET exon 14 skipping mutation patients with locally advanced or metastatic non-small-cell lung cancer (NSCLC).Methods/designIn this two-part multicenter phase Ib study, eligible patients with locally advanced or metastatic NSCLC harboring c-MET overexpression or MET exon 14 skipping mutation were enrolled into Part A (tested positive for c-MET overexpression [immunohistochemical staining score ≥ 2+]; 300 mg quaque die [QD], 450 mg QD and 600 mg QD cohorts) or Part B (tested positive for MET exon 14 skipping mutation; 400 mg bis in die [BID] cohort), respectively. The primary endpoints were safety, objective response rate (ORR) and disease control rate (DCR), the second endpoints were PK parameters, progression-free survival (PFS) and overall survival (OS).ResultsBetween March 15, 2017 and September 18, 2021, 38 patients were enrolled (Part A, n = 34; Part B, n = 4). Of 38 patients, 32 (84.2%) patients completed the treatment protocol. As of the data cut-off date on January 27, 2022, all patients reported at least one treatment-emergent adverse event (TEAE). Ninety-two point one percent (35/38) of patients experienced treatment-related adverse events (TRAEs), and grade ≥ 3 TRAEs were observed in 11 (28.9%) patients. The most common TRAEs were elevated alanine aminotransferase (ALT, 14/38, 36.8%) and elevated aspartate aminotransferase (AST, 11/38, 28.9%). Only one (2.6%) patient had treatment-related serious adverse event (SAE) in 600 mg QD cohort due to thrombocytopenia. PK analysis showed BPI-9016M and its main metabolites (M1 and M2-2) reached steady state after seven days of continuous administration. At the dose of 300 mg QD and 450 mg QD, the exposure of BPI-9016M increased with increasing dose. Exposure of BPI-9016M was similar at 450 mg QD and 600 mg QD, which may exhibit a saturation trend. In all patients, ORR and DCR were 2.6% (1/38, 95% confidence interval [CI] 0.1–13.8%) and 42.1% (16/38, 95% CI 26.3–59.2%), respectively. Only one partial response (PR) patient was observed at a dose of 600 mg QD in Part A. In Part B, DCR was 75.0% (3/4, 95% CI 19.4–99.4%). The median PFS and OS in all 38 patients were 1.9 months (95% CI 1.9–3.7) and 10.3 months (95% CI 7.3–not evaluable [NE]), respectively.ConclusionBPI-9016M showed manageable safety profile in c-MET overexpression or MET exon 14 skipping mutation patients with locally advanced or metastatic NSCLC, but showed limited efficacy.Trial registrationClinicaltrials.gov NCT02929290 (11/10/2016).

Abstract 847: Applying the principles of convexity and concavity to guide treatment scheduling of ALK inhibitors in non-small cell lung cancer

Abstract Recently, the US Food and Drug Administration (FDA) has initiated “Project Optimus,” designed to revisit and reform the decades-old maximum-tolerable dose paradigm. Traditional dose selection schemes designed for cytotoxic therapies may not apply for targeted therapies with exposure-response curves that often plateau below maximum tolerable patient toxicity levels. While nonlinear sigmoidal exposure-response curves are ubiquitous in oncology, they are typically used to measure differential response in first-order effects (mean value of drug dose delivered), while second-order effects (variance of drug dose) are generally ignored. Herein we present a paradigm for guiding treatment scheduling based on the convexity (or its converse, concavity) of exposure-response curves - a phenomenon that is widely applicable to targeted therapies. The “curvature” of exposure-response curves (e.g. a convex or concave shape) provides a direct prediction of continuous treatment, compared with high-dose/low-dose treatment. For example, if the exposure-response function is concave near a dose of ‘x’, continuous (daily) administration of x may be less effective response compared to a regimen that switches equally between 120% of x and 80% of x (every other day), even though both regimens use the same total drug. This paradigm is specifically relevant to targeted therapies which, as stated above, are associated with exposure-response plateaus that are concave in nature (to be compared with convex exposure-response curves common in cytotoxic chemotherapies). Analysis of response curves in vitro for a H3122 ALK-positive non-small cell lung cancer (NSCLC) cell line predicts that evolved-resistance lines are generally more concave, while treatment-naïve lines are more convex. In vivo, selection pressure due to treatment selects for resistant phenotypes over time. Previous literature shows resistance to ALK inhibitors occurs gradually, as tumors acquire cooperating genetic and epigenetic adaptive changes. Thus, we hypothesized the existence of a critical point in the time-evolution of ALK-positive tumors where it is optimal to switch from continuous treatment to high-dose/low-dose to mitigate the onset of gradual resistance. In this work, we construct a mathematical modeling framework of gradual resistance, parameterized to data, and predict time-dependent curvature in continuous (8 weeks), volatile (8 weeks) ALK inhibition in vivo. our key insight is that curvature increases in proportion to the amount of resistance in the tumor population. Curvature provides a time-dependent metric which 1) predicts the emergence of resistance and 2) determines the optimal subsequent dosing strategy. We test our key hypothesis in vivo, comparing continuous and volatile treatment schedules of ALK inhibitors to a switching schedule of continuous-volatile (4 weeks each). Citation Format: Jeffrey West, Bina Desai, Maximilian Strobl, Luke Pierik, Robert Vander Velde, Mark Robertson-Tessi, Andriy Marusyk, Alexander R. Anderson. Applying the principles of convexity and concavity to guide treatment scheduling of ALK inhibitors in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 847.

Metabolic Disposition and Elimination of Tritum-Labeled Sulfamethoxazole in Pigs, Chickens and Rats.

Sulfamethoxazole (SMZ), as a sulfa antibiotic, is often used in the treatment of various infectious diseases in animal husbandry. At present, SMZ still has many unresolved problems in the material balance, metabolic pathways, and residual target tissues in food animals. Therefore, in order to solve these problems, the metabolism, distribution, and elimination of SMZ is investigated in pigs, chickens, and rats by radioactive tracing methods, and the residue marker and target tissue of SMZ in food animals were determined, providing a reliable basis for food safety. After a single administration of [3H]-SMZ (rats and pigs by intramuscular injection and chickens by oral gavage), the total radioactivity was rapidly excreted, with more than 93% of the dose excreted within 14 days in the three species. Pigs and rats had more than 75% of the administered volume recovered by urine. After 7 days of continuous administration, within the first 6 h, radioactivity was found in almost all tissues. The highest radioactivity and longest persistence in pigs was in the liver, while in chickens it was in the liver and kidneys, most of which was removed within 14 days. A total of six, three and three metabolites were found in chickens, rats and pigs, respectively. N4-acetyl-sulfamethoxazole (S1) was the main metabolite of SMZ in rats, pigs and chickens. The radioactive substance with the longest elimination half-life is sulfamethoxazole (S0), so S0 was suggested to be the marker residue in pigs and chickens.

Salvia miltiorrhiza polysaccharides alleviate florfenicol-induced inflammation and oxidative stress in chick livers by regulating phagosome signaling pathway

Florfenicol (FFC) is a commonly used antibiotic in animal breeding, especially in broiler breeding. Previous studies found that FFC could affect the liver function of chickens. However, the mechanisms underlying the effects of FFC on liver function are still not completely clear. Moreover, the research on drugs that antagonize FFC hepatotoxicity is relatively lacking. Salvia miltiorrhiza polysaccharides (SMPs) have been proved to have obvious liver protection effects. Therefore, we exposed chicks to FFC at the clinically recommended dose of 0.15 g/L. At the same time, 0.15 g/L FFC and 5 g/L SMPs were given to another group of chicks. After 5 days of continuous administration, the livers of chicks from different treatment groups were sequenced by transcriptome and proteome. Based on the analysis of sequencing data, we also focused on the detection of inflammation and oxidation indicators related to the phagosome signaling pathway with significant enrichment of differential factors in the livers of chicks. The results showed that some significantly differentially expressed genes and proteins induced by FFC were enriched in the phagosome signaling pathway, and they increased the expression levels of inflammatory factors and peroxides. However, SMPs intervention significantly reversed the tendency of FFC to alter phagosome signaling pathways and reduced the expression levels of inflammatory factors and peroxides. In conclusion, FFC caused liver inflammation and oxidative stress in chicks by regulating the phagosome signaling pathway. Meanwhile, SMPs could improve the adverse effects of FFC on the phagosome signaling pathway. This study provided new insights into the ameliorative effects and mechanisms of SMPs on hepatotoxicity of FFC.

Efficacy of Bacillus coagulans BC01 on loperamide hydrochloride-induced constipation model in Kunming mice.

In this study, the laxative effect of Bacillus coagulans BC01 (BC01) in mice was investigated using a functional constipation mouse model. Six-week-old male specific pathogen-free (SPF) Kunming mice were randomly divided into five groups: normal control group (saline), model group (loperamide hydrochloride), drug control group (bisacodyl), BC01 low-dose group (4.0 × 108 CFU/mL) and BC01 high-dose group (4.0 × 109 CFU/mL). Except for the normal group, the functional constipation model was established by administering 0.25 mL of a loperamide hydrochloride suspension (1 mg/mL) twice daily for four consecutive days by oral gavage. After modeling, the BC01 groups were administered 0.25 mL of BC01. The bisacodyl served as a control and was administered orally at a dose of 100 mg/kg, while the other groups were administered 0.25 mL of sterile saline. After 7 days of continuous administration, the experimental mice were again induced by loperamide hydrochloride. During this period, the mechanism of BC01 to improve constipation symptoms in mice was analyzed by measuring the changes in body weight, fecal water content, small intestine propulsion rate, histology of small intestinal tissue sections, fecal microbial diversity, serum indices, as well as mRNA and protein expression levels in the small intestinal tissue. BC01 was found to significantly promote the intestinal propulsion rate and increase the fecal water content in the mice. BC01 could also alleviates constipation by regulating gastrointestinal motility (substance P, motilin, endothelin-1, somatostatin, and vasoactive intestinal peptide), gene expression (c-Kit, SCF, COX-2, NF-κB, iNOS, and eNOS), intestinal inflammation (eNOS, iNOS, NF-κB), and the intestinal microbiota composition in the constipated mice. In addition, the high-dose BC01 treatment had the best preventive effect on constipation. BC01 is a probiotic strain to effectively relieve the adverse effects of constipation.

Chloral Hydrate Alters Brain Activation Induced by Methamphetamine-Associated Cue and Prevents Relapse.

Methamphetamine is a highly addictive drug and its abuse leads to serious health and social problems. Until now, no effective medications are yet available for the treatment of methamphetamine addiction. Our study reveals that chloral hydrate, a clinical sedative drug, suppresses the seeking desire for methamphetamine. After 5 days of continuous administration (subanesthetic dose 50 mg/kg and 100 mg/kg), methamphetamine-seeking behavior of rats was inhibited in the condition place preference and intravenous self-administration tests. Furthermore, chloral hydrate treatment robustly suppressed cue-induced methamphetamine relapse. The whole brain c-fos immunostaining revealed that chloral hydrate treatment suppressed neuronal activity in the rhomboid thalamic nucleus (Rh), dorsal endopiriform nucleus (dEn), and claustrum (Cl) while enhanced zona incerta (ZI) activity during cue-induced methamphetamine relapse. Therefore, chloral hydrate could remodel neural network activity and serve as a potential medicine to treat methamphetamine addiction.