Middle Dose Research Articles

HUC-MSC transplantation therapy effects on lupus-prone MRL/lpr mice at early disease stages

BackgroundThe efficacy of human umbilical cord mesenchymal stem cell (hUC-MSC) transplantation in treating systemic lupus erythematosus (SLE) has been confirmed by small-scale clinical trials. However, these trials focused on severe or refractory SLE, while few studies focused on mild SLE. Therefore, this study focused on the therapeutic effects of hUC-MSC transplantation in early-stage or mild MRL/lpr lupus model mice.MethodsCommercially available hUC-MSCs were transplanted into 8-week-old MRL/lpr mice by tail vein injection. Flow cytometry was used to analyze B cells and their subsets in the peripheral blood. Further, plasma inflammatory factors, autoantibodies, and plasma biochemical indices were detected using protein chip technology and ELISA kits. In addition, pathological staining and immunofluorescence were performed to detect kidney injury in mice.ResultshUC-MSC transplantation did not affect the mice’s body weight, and both middle and high dose hUC-MSC transplantation (MD and HD group) actually reduced spleen weight. hUC-MSC transplantation significantly decreased the proportion of plasmablasts (PB), IgG1− PB, IgG1+ PB, IgG1+ memory B (MB) cells, IgG1+ DN MB, and IgG1+ SP MB cells. The hUC-MSC transplantation had significantly reduced plasma levels of inflammatory factors, such as TNF-α, IFN-γ, IL-6, and IL-13. Pathological staining showed that the infiltration of glomerular inflammatory cells was significantly reduced and that the level of glomerular fibrosis was significantly alleviated in hUC-MSC-transplanted mice. Immunofluorescence assays showed that the deposition of IgG and IgM antibodies in the kidneys of hUC-MSC-transplanted mice was significantly lower than in the control.ConclusionhUC-MSC transplantation could inhibit the proliferation and differentiation of peripheral blood B cells in the early-stage of MRL/lpr mice, thereby alleviating the plasma inflammatory environment in mice, leading to kidney injury remission. The study provides a new and feasible strategy for SLE treatment.

Amelioration of oxidative kidney damage in offspring by maternal trans-fatty acid exposure in mice by secoisolariciresinol diglucoside.

Trans-fatty acids (TFAs), primarily derived from the food industry's production processes, have become a globally recognized public health issue due to the detrimental impact they have on human well-being. Secoisolariciresinol diglucoside (SDG) is a polyphenolic compound derived from flax lignans, possessing antioxidative properties. This study aims to investigate the protective effect of SDG on kidney oxidative damage in offspring of mice caused by maternal exposure to TFA during pregnancy and lactation. A total of 30 c57BL/6 female rats were randomly divided into 5 groups: a control group, a TFA-exposed group, a low-(TFA+LSDG) group, a medium-(TFA+MSDG) group, and a high-(TFA+HSDG) group (n=6 in each group). With the exception of the control group, the maternal mice in the remaining 4 groups received a daily oral gavage of TFA at a dosage of 60 mg/(kg·BW) throughout the experimental period. The mothers in the control group were administered physiological saline via oral gavage once daily. Meanwhile, the 3 SDG intervention groups were provided with ad libitum access to SDG feed containing 10 mg/kg (low), 20 mg/kg (medium), and 30 mg/kg (high) of SDG. The female mice were conceived overnight. If the vaginal plug appeared in the next morning, the female mice were conceived and included in the experimental stage until the end of the 21th day lactation period. The body weight and kidney mass of offspring were recorded, and the kidney coefficient was calculated. The kidney was detected by HE staining to observe the histopathological changes, and the level of reactive oxidative species (ROS) was detected by fluorescence probe-dihydroethidium (DHE) staining; the expression levels of total superoxide dismutase (T-SOD) and malondialdehyde (MDA) in renal homogenate were detected, and the expression of nuclear factor E2-related fator2 (Nrf2) and hemeoxygenase-1 (HO-1) protein was analyzed by immunohistochemistry (IHC) staining. The mRNA expressions of Nrf2 and HO-1 were detected by real-time PCR, and the protein expression of Cu/Zn-superoxide dismutase (Cu/Zn-SOD), Mn-superoxide dismutase (Mn-SOD), glutathione peroxidase-1 (GPx-1), Nrf2 and HO-1 were detected by Western blotting. Compared with the control group, the kidney coefficient in the TFA-exposed group was increased, the morphology and structure of kidney tissue was abnormal; the activity of T-SOD enzyme was decreased, and the content of MDA was increased, the level of ROS was increased; the expressions of Cu/Zn-SOD, Mn-SOD, GPx1 protein were decreased, and the mRNA and protein expressions of Nrf2 and HO-1 were decreased, there were all significant difference (all P<0.05). Compared with the TFA-exposed group, the ROS levels were reduced, and the T-SOD enzyme activity as well as the protein expression of Cu/Zn-SOD, GPx-1, Mn-SOD, Nrf2 and HO-1 were up-regulated in the low, middle and high dose SDG intervention groups; the kidney coefficient and MDA content were decreased in the middle and high dose SDG groups; the Nrf2 mRNA expression in the high dose SDG group was up-regulated, there were all significant difference (all P<0.05). Maternal exposure to TFA during pregnancy and lactation can lead to oxidative damage in the kidney of offspring, and the SDG intervention may alleviate TFA-induced oxidative damage by up-regulating the expression of Nrf2 and HO-1 signal pathway.

Houpo Paiqi Mixture Promotes Intestinal Motility in Constipated Rats by Modulating Gut Microbiota and Activating 5-HT-cAMP-PKA Signal Pathway.

Constipation is a common functional gastrointestinal disorder which needs more effective treatment approaches. Houpo Paiqi Mixture (HPPQM) is a type of Chinese patent medicine developed from a classical formula that has been widely applied to the treatment of intestinal motility disorder. Here we aim to assess the effectiveness of HPPQM in the treatment of constipation in rat models and its potential mechanism. UPLC-MS/MS was performed to investigate the chemical component of HPPQM. Rats were randomly divided into normal control, constipation model (CM), HPPQM (low, middle and high dose) and mosapride groups. Loperamide 8mg/kg was given orally to induce constipation model. The small intestine motility, colonic contraction, rectum propulsion and histological feature of the colon were significantly improved in HPPQM group, compared with CM group (P<0.05). Results of 16S rRNA sequencing revealed that HPPQM treatment strikingly restructured intestinal microbiota in constipated rats by increasing the relative abundances of Bacteroides and Akkermansia and decreasing the relative abundances of Prevotella and Lactobacillus. The levels of GPR43, 5-HT, 5-HT4R, cAMP, PKA were decreased while SERT was increased in constipated rats (P<0.05), which could be restored to normal levels by treatment with HPPQM (P<0.05). Differences in amplitude between experimental CLSMs (with HPPQM added) and control CLSMs were discovered, starting at the concentration of 40 nL/mL (P<0.05). It was found that GLPG0974 and GR113808 could significantly reduce this reactivity (P<0.05). HPPQM manifested a curative effect in constipated rats by promoting intestinal motility. The underlying mechanisms might be related to modulating gut microbiota and activating 5-HT-cAMP-PKA signal pathway.

Transcriptomic and metabolomic insights into the roles of exogenous β-hydroxybutyrate acid for the development of rumen epithelium in young goats

Beta-hydroxybutyric acid (BHBA), as one of the main metabolic ketones in the rumen epithelium, plays critical roles in cellular growth and metabolism. The ketogenic capacity is associated with the maturation of rumen in young ruminants, and the exogenous BHBA in diet may promote the rumen development. However, the effects of exogenous BHBA on rumen remain unknown. This is the first study to investigate the mechanisms of BHBA on gene expression and metabolism of rumen epithelium using young goats as a model through multi-omics techniques. Thirty-two young goats were divided into control, low dose, middle dose, and high dose groups by supplementation of BHBA in starter (0, 3, 6, and 9 g/day, respectively). Results demonstrated the dietary of BHBA promoted the growth performance of young goats and increased width and length of the rumen papilla (P < 0.05). Hub genes in host transcriptome that were positively related to rumen characteristics and BHBA concentration were identified. Several upregulated hub genes including NDUFC1, NDUFB4, NDUFB10, NDUFA11 and NDUFA1 were enriched in the gene ontology (GO) pathway of nicotinamide adenine dinucleotide (NADH) dehydrogenase (ubiquinone) activity, while ATP5ME, ATP5PO and ATP5PF were associated with ATP synthesis. RT-PCR revealed the expression of genes (HMGCS2, BDH1, SLC16A3, etc.) associated with lipolysis increased significantly by BHBA supplementation (P < 0.05). Metabolomics indicated that some metabolites such as glucose, palmitic acid, cortisol and capric acid were also increased (P < 0.05). This study revealed that BHBA promoted rumen development through altering NADH balance and accelerating lipid metabolism, which provides a theoretical guidance for the strategies of gastrointestinal health and development of young ruminants.

Yinchenhao Tang alleviates high fat diet induced NAFLD by increasing NR1H4 and APOA1 expression.

Traditional Chinese medicine Yinchenhao Tang (YCHT) demonstrated benefits when treating nonalcoholic fatty liver disease (NAFLD), but the dose effects and potential targets are still ambiguous. In this study, different concentrations of YCHT were employed to treat NAFLD and the underlying therapeutic targets were investigated. Kunming mice were fed with high fat diet (HFD) for 8 weeks to induce NAFLD, then treated with 3 different concentrations of YCHT. Hepatic pathological changes and serum lipid levels were examined. Network pharmacology was applied to screen the potential targets of YCHT for NAFLD modulation. NR1H4 and APOA1 expression was evaluated by QPCR and western blotting. Immunohistochemistry (IHC) staining was conducted to visualize the localization pattern of NR1H4 and APOA1 in the liver. YCHT significantly reduced liver lipid storage and improved the liver pathological status of NAFLD mice. The serum lipid levels, as well as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, were remarkably reduced by the middle and high dose YCHT. There are 35 potential targets for YCHT to regulate NAFLD. HFD suppressed both RNA and protein expression of NR1H4 and APOA1, while YCHT elevated NR1H4 and APOA1 expression. IHC staining indicated that NR1H4 was mainly located in the cell nucleus and the APOA1 signal was observed at the liver sinusoid or cytoplasm. YCHT can effectively ameliorate HFD induced NAFLD by modulating the promising targets of NR1H4 and APOA1.

Safety profile of colocasia esculenta tuber extracts in benign prostate hyperplasia

IntroductionThis study was motivated by the increasing global incidence of benign prostatic hyperplasia (BPH) and the promising potential of nutraceuticals as complementary therapies in ameliorating its burden. We report the safety profile of C. esculenta tuber extracts, a novel nutraceutical in benign prostate hyperplasia in a rat model.MethodsIn this study, forty-five male albino rats were randomly assigned to 9 groups of 5 rats each. Group 1 (normal control) received olive oil and normal saline. Group 2 (BPH untreated group) received 3 mg/kg of testosterone propionate (TP) and normal saline, and group 3 (positive control) received 3 mg/kg of TP and 5 mg/kg of finasteride. Treatment groups 4, 5, 6, 7, 8, and 9 received 3 mg/kg of TP and a middle dose (200 mg/kg) of LD50 of ethanol crude tuber extract of C. esculenta (ECTECE) or hexane, dichloromethane, butanone, ethyl acetate and aqueous fractions of ECTECE respectively for a period of 28 days.ResultsThe negative controls showed a significant (p < 0.05) increase in mean relative prostate weight (approximately 5 times) as well as a reduction in relative testes weight (approximately 1.4 times less). There was no significant (p > 0.05) difference in the mean relative weights of most vital organs: liver, kidneys, and heart. This was also observed in hematological parameters: RBC, hemoglobin, HCT, MCV, MCH, MCHC, and platelets counts. In general, we note that the effects of the well-established drug finasteride on the biochemical parameters and histology of selected organs are comparable to those of C. esculenta fractions.ConclusionThis study demonstrates that C. esculenta tuber extracts provide potentially safe nutraceutical if applied in the management of benign prostate hyperplasia based on a rat model.

EpCAM CAR T (IMC001) for the treatment of advanced GI cancers.

4034 Background: EpCAM is highly expressed in various cancer types of the GI system and at metastatic sites. It serves as a promising prognostic/predictive marker (CTC) and therapeutic target. IMC001 is an EpCAM-targeted CAR-T cell that showed promising anti-tumor activities in preclinical studies. Here, we report the results of the low and middle dosage groups in advanced gastric cancers (CT03) and GI cancers (CT04). Methods: This first-in-human, open-label trial involved two separate single-site trials. Both followed a classic 3+3 design with dose escalation of 0.3, 1 or 3 million CAR-T cells/kg after lymphodepletion chemotherapy. CT03 was an IMC001 monotherapy trial (Stage 1) for gastric cancer, while CT04 had Stage 1 and the combination with RFA or microwave ablation (Stage 2) for GI cancers. Eligible patients were those with EpCAM-positive (more than 10%) cancers who had no further standard treatment options and were ECOG 0 or 1. The objective was to assess the safety, PK/PD profile and preliminary efficacy of IMC001. Results: As of January 30, 2023, 12 patients with 6 colorectal and 6 gastric cancers had enrolled, with half receiving 0.3 million and the other half receiving 1 million cells/kg IMC001 infusion. No patient experienced DLT within the 4-week follow-up visits after infusion. All patients experienced ≥Grade 3 hematologic toxicity. One patient in the low-dose group had a SAE of immune hepatitis (Grade 3), which might have been related to cell therapy, and occurred around 11 days after the CAR-T infusion, prolonging the patient's hospitalization. Manageable CRS (Grade 1 to 3) and no ICANS were observed. Other adverse events related to cell therapy were CTCAE Stage 1-2 nausea, vomiting, asthenia, or pruritus, and these recovered quickly. Analyses of the CAR-T cells in peripheral blood revealed robust engraftment in all patients, with the peak number of CAR+ cells reaching on day 5-7 after infusion. CTC remained under the detection limit for more than 40 weeks after cell infusion. Significant elevations of serum levels of IL-6, IP-10, IFN-γ, IL-15, and MCP-1 were observed in most patients. Preliminary efficacy data showed that 2 out of 6 advanced gastric cancer patients were evaluated as PR and 3 remained SD by RECIST 1.1 at the low and middle dosages (CT03). The first PR patient received a second IMC001 infusion on week 50 and had survived for more than 60 weeks. The second PR patient underwent successful surgical removal of the stomach 28 weeks after IMC001 infusion. The CT04 trial is ongoing. Conclusions: This is the first CAR-T therapy ever tested in humans targeting the EpCAM. IMC001 showed a favorable safety profile and reasonable anti-tumor activities at the low and middle dosage levels in patients with advanced EpCAM+ GI cancers, especially in gastric cancer. In addition, our trial has successfully provided a surgical treatment opportunity after CAR-T therapy downstaging of unresectable gastric cancers. Updated data from open cohorts will be presented. Clinical trial information: NCT05028933 .

Clinical-grade natural killer cells expanded from donor peripheral blood and anti-leukemia effect against myeloid leukemia.

e19018 Background: Natural killer (NK) cells play a crucial role in immune surveillance and tumor clearance. It has been reported that allogeneic NK cell therapy has good safety and curative effect in patients with refractory and relapsed acute myeloid leukemia (AML). However, the clinical application of adoptive NK infusion is limited due to the lower number of human NK cells. We established a feeder-free NK cell expansion system from peripheral blood (PB) and explore the anti-leukemia potential in preclinical and clinical studies. Methods: The killing capacity of ex vivo-expanded NK cells were detected by Flow cytometry. The in vivo expansion, survival and anti-leukemia activity of the expanded NK cells were detected in K562-bearing NSG mice. Six AML patients with hematological/extra medullary relapsed (n = 3) or persistent positive minimal residual disease (n = 3) after consolidation and maintenance therapy were enrolled in Peking University Institute of Hematology from 2021 to 2022. The patients were treated with high-dose cyclophosphamide and fludarabine, then combined with adoptive infusion of donor-derived expanded NK cells. The chimerism, the number and function of NK cells were analyzed, and the safety and clinical efficacy were evaluated. Results: The ex vivo-expanded NK cells exhibit highly anti-leukemia activity. NK cells proliferate in mice and the CD56+ cells to reach the peak time in each organ is 3-7 days and drop to the level before infusion after 14 days. NK cell infusion significantly improve the survival of leukemia mice, and the anti-leukemia effect is NK cell dose-dependent. In the clinical trial, six patients received a total of 7 cases of NK cell infusion, including 3 cases in low-dose group and 4 cases in middle-dose group, among which 1 patient received 2 dose (1 low-dose and 1 middle-dose). In the low-dose group, 1 patient got response (1/3), with the minimal residual disease turned negative. In the middle-dose group, 3 patients responded (3/4), of which 1 patient had the fusion gene CBFB /ABL decreased from 0.99% to 0.13%, 1 case of extramedullary recurrence of AML turned negative, and NK cells could be detected in cerebrospinal fluid, 1 case of hematological recurrence got hematological remission after treatment. The survival time of NK cells in vivo was 7d-30d, with longer time in middle dose group than those in low dose group. The proportion and number of NK cells in responding cases were significantly higher than those in non-responsive group. Conclusions: Clinical grade expanded NK cells has strong anti-leukemia activity in patients with relapse or minimal residual positive AML patients. NK cells can survive for 2 weeks and can enter the central nervous system without obvious adverse reactions. The response rate in middle-dose group is higher than that in low-dose group. However, the efficacy of NK cells still needs to be further confirmed in a large sample size. Clinical trial information: NCT04221971 .

Chlorophyll Content and Leaf Morphological Features of Capsicum annum L. cv. ‘Alexander’ under Yeast and Caffeine Tested as Bio-fertilizers

Bio-fertilizers are in the present more and more of interest as alternative nutritive inputs. This research aim was to test different doses of yeast and caffeine on pepper growth and chlorophyll content. The experiment in controlled conditions consisted of 8 phytocosms treatments with 3 yeast and caffeine doses 1%, 2%, and 3%, one mixed treatment (2%), and the control treatment in five replications. The yeast addition promoted all the tested morphological parameters especially the middle dose tested. Caffeine treatments induced a slow growth of pepper vegetative period, even a delayed development compared with the control treatment. In conclusion, the mixed treatment was the most efficient pepper bio-fertilizer and provided the highest values of leaf length, width, seedling height, and also total chlorophyll content.

The Role of G3BP1 Gene Mediates P38 MAPK/JNK Pathway in Testicular Spermatogenic Dysfunction Caused by Cyfluthrin.

In recent years, male infertility has received global attention and seriously affected the quality of human fertility, and pyrethroids (type II pyrethroids), as recognized environmental endocrine disruptors, may threaten male reproductive health. Therefore, in this study, we established an in vivo model for the development of testicular and germ cell toxicity induced by cyfluthrin and explored the role and mechanism of the G3BP1 gene-mediated P38 MAPK/JNK pathway in testicular and germ cell damage caused by cyfluthrin to find early and sensitive indicators and new therapeutic targets for the development of testicular damage. Firstly, 40 male Wistar rats (about 260 g) were divided into a control group (corn oil), low dose group (6.25 mg/kg), middle dose group (12.5 mg/kg) and high dose group (25 mg/kg). The rats were anesthetized and executed after 28 days of poisoning on alternate days. Then, HE staining, transmission electron microscopy, ELISA, q-PCR, Western blot, immunohistochemistry, double-immunofluorescence and TUNEL were used to observe the pathology, androgen levels, oxidative damage and altered expression of the key factors of the G3BP1 and MAPK pathways in rat testes. The results showed that, compared with the control group, the testicular tissue and spermatocytes were superficially damaged with an increasing dose of cyfluthrin; furthermore, it could interfere with the normal secretion of the hypothalamic-pituitary-gonadal axis (serum GnRH, FSH, T and LH levels) and cause hypergonadal dysfunction. A dose-dependent increase in MDA and a dose-dependent decrease in T-AOC indicated that the oxidative-antioxidative homeostatic balance was disrupted. The Western blot and qPCR analysis revealed that G3BP1, p-JNK1/2/3, P38 MAPK, p-ERK, COX1 and COX4 proteins and mRNA expression were decreased, and p-JNK1/2/3, p-P38MAPK, caspase 3/8/9 proteins and mRNA expression were significantly increased. The double-immunofluorescence and immunohistochemistry results showed that the protein expression of G3BP1 decreased with an increasing dose of staining, while the expression of JNK1/2/3 and P38 MAPK were increased significantly. The positive expressions of G3BP1 were mainly located in the testicular germinal epithelium and germ cell layer, and the positive expressions of JNK1/2/3 were mainly located in the testicular germinal epithelium and sperm cells, while the positive expressions of P38 MAPK were located in all levels of the germ cells and spermatozoa. Our results demonstrated that exposure to cyfluthrin caused testicular and spermatocyte damage in rats, which could cause pathomorphology, altered androgen levels and a decreased antioxidant capacity. When the intracellular antioxidant capacity was impaired, G3BP1 expression and activity were inhibited, causing activation of the P38 MAPK/JNK pathway and activation of the intracellular apoptotic pathway, which, in turn, led to germ cell apoptosis.

Polyethylene Microplastic Particles Alter the Nature, Bacterial Community and Metabolite Profile of Reed Rhizosphere Soils

With the wide use of polyethylene film, the influence of polyethylene microplastic particles produced by its weathering on the rhizosphere soil microenvironment has attracted more and more attention from scientific research circles. In this study, the effects of low (0.2% w/w), medium (1% w/w), and high (2% w/w) doses of polyethylene particles and the combined reed biomass (2% w/w) on soil environmental factors and bacterial communities and metabolites in the reed rhizosphere were evaluated by a 90-day pot microscopic simulation system. The shape and surface microstructure of polyethylene particles in each treatment group changed obviously. A high (2% w/w) dose of microplastics significantly increased the TKN, TOC, and TP in reed root soil. The addition of the biomass significantly improved the activities of urease and sucrase in the soil. The α diversity of bacteria was not significantly affected by the addition of LDPE microplastics and biomass, but the β diversity of the bacterial community and the relative abundance of the Candidatus_Roku Bacteria, Chloroflexi, Unclassified_Blastocatella_Genus were significantly changed by the addition of middle (1% w/w) and high (2% w/w) doses of microplastics. In addition, the spectrum analysis of the soil metabolites showed that the abundance of soil metabolites was changed in each treatment group, and the differential metabolites were significantly up-regulated or down-regulated. Our findings provide a scientific reference to elucidate the impact of LDPE microplastic particles on the inter-rooted soil microenvironment and improve our understanding of the potential risks of microplastics in soil ecosystems.

Pengaruh Pemberian Kombinasi Ekstrak Kulit Pisang dan Kulit Nanas terhadap Indeks Organ Tikus Wistar

Banana peel and pineapple peel contain potassium which may be developed into health supplements. The development of banana peels and pineapple peels as ingredients for potassium supplements must meet safety aspects that can be proven by subchronic toxicity tests. This subchronic toxicity test aimed to determine the effect of repeated administration of a combination of banana peel and pineapple peel extract for 28 days on the liver, kidney, kidney, lung, heart, and spleen index of Wistar rats. This research used male and female white rats which were divided into six treatment groups, namely the control group, low dose, middle dose, high dose, satellite control, and high dose satellite. The results showed that there was a significant increase in the liver and kidney organ index, and a significant decrease in the adrenal gland organ index after administration of a combination of banana peel and pineapple peel extract. This research concludes that the combination of banana peel and pineapple peel extract has an effect on the liver, kidney, and adrenal gland organ index in Wistar white rats.

Comparison between supine and prone patient setup for lumbosacral spinal stereotactic body radiosurgery with CyberKnife.

The aim of this study is to analyze which tracking modality is more suitable for stereotactic body radiosurgery of lumbosacral spinal tumors by comparing prone and supine patient treatment setup. Eighteen patients with lumbosacral spinal tumors were selected. CT simulation was performed in the supine position (fixed with a vacuum cushion) and prone position (fixed with a thermoplastic mask and prone plate), respectively. The plans in the supine and prone positions were designed using the xsight spine tracking (XST) and xsight spine prone tracking (XSPT) modalities, respectively. The dose-volume histogram (DVH) parameters, namely, V100%, D95%, Dmean, conformity index (CI), and heterogeneity index (HI) in planning target volume (PTV), as well as Dmax, D0.1cc, D1cc, and D5cc in the cauda equina and bowel were recorded. The supine plans were simulation plans and were not used for treatment, which were only used to record the alignment errors. The spinal tracking correction errors (alignment error) and correlation errors of the synchrony respiratory model in the prone position were recorded during the treatment. After treatment, the simulation plan of the supine position was implemented and the spinal tracking correction errors were recorded. The parameters of correction error and DVH parameters for the two positions were analyzed using the paired t-test to compare the difference in positioning accuracy and dose distribution. In addition, the correlation errors of the synchrony respiratory model in the prone position were analyzed to evaluate the prediction accuracy of the synchrony model. For patient setup, the correction error of the supine position in interior/posterior was (0.18 ± 0.16) mm and the prone position was (0.31 ± 0.26) mm (P< 0.05). The correction error of the supine position in inferior/superior was (0.27 ± 0.24) mm, and the prone position was (0.5 ± 0.4) mm (P< 0.05). The average correlation errors of the synchrony model for left/right, inferior/superior, and anterior/posterior in the prone position were (0.21 ± 0.11) mm, (0.41 ± 0.38) mm, and (0.68 ± 0.42) mm, respectively. For the dose distribution, compared with prone plans, the average CI in supine plans was increased by 4.5% (P< 0.05). There was no significant difference in HI, PTV V100%, D95%, and Dmean between the prone and supine plans. Compared with supine plans, average D1cc and D5cc for the cauda equina was significantly decreased by 4.7 and 15.3% in the prone plan (P< 0.05). For the bowel, average Dmax, D0.1cc, D1cc, and D5cc were reduced by 8.0, 7.7, 5.2, and 26.6% in prone plans (P< 0.05) compared with supine plans. Compared with the supine setup, the prone setup combined with XSPT modality for the lumbosacral spinal stereotactic body radiosurgery can spare the bowel and cauda equina of the middle and low dose irradiation, and decrease the number of beams and monitor units.

Effects of Acetaminophen on Reproductive Activities in Male Golden Hamsters

Acetaminophen [Paracetamol, N-acetyl-para-aminophenol (APAP)] is a common over-the- counter analgesic agent as nonsteroidal anti-inflammatory drugs (NSAIDs). The high doses or the long-term treatment of acetaminophen via usual gavage feeding resulted in damage of testicles that presented recoverable impairment, as well as liver and kidney. The influence of acetaminophen was examined in male golden hamsters treated with acetaminophen- containing diet feeding. They were divided into 5 groups and subjected to this experiment for 4 weeks: animals housed in long photoperiod (LP) as LP control, animals housed in short photoperiod (SP) for 4 weeks as SP control (SP4), and groups of animals treated with low, middle, and high concentrations of acetaminophen (Low, Middle, High groups). Also animals housed in SP for 8 weeks were included (SP8) to contrast testicular activities, if necessary. As results, spermatozoa filled the seminiferous tubules of the testicles of animals in LP control and SP4 groups. The aspects were seen in the animals taken diets of low and middle doses of acetaminophen. The animals who fed high dose of acetaminophen showed large or small testicles. The large testicles displayed all germ cells at the steps of spermatogenesis. The small testicles presented no sperm as the animals housed in SP for 8 weeks. Thus these results indicate that acetaminophen invokes the antigonadal effects and accelerates the regressing process of the testicles in the animals compared to the animals exposed to SP.

Effects of Acetaminophen on Reproductive Activities in Male Golden Hamsters

Acetaminophen [Paracetamol, N-acetyl-para-aminophenol (APAP)] is a common over-the- counter analgesic agent as nonsteroidal anti-inflammatory drugs (NSAIDs). The high doses or the long-term treatment of acetaminophen via usual gavage feeding resulted in damage of testicles that presented recoverable impairment, as well as liver and kidney. The influence of acetaminophen was examined in male golden hamsters treated with acetaminophen- containing diet feeding. They were divided into 5 groups and subjected to this experiment for 4 weeks: animals housed in long photoperiod (LP) as LP control, animals housed in short photoperiod (SP) for 4 weeks as SP control (SP4), and groups of animals treated with low, middle, and high concentrations of acetaminophen (Low, Middle, High groups). Also animals housed in SP for 8 weeks were included (SP8) to contrast testicular activities, if necessary. As results, spermatozoa filled the seminiferous tubules of the testicles of animals in LP control and SP4 groups. The aspects were seen in the animals taken diets of low and middle doses of acetaminophen. The animals who fed high dose of acetaminophen showed large or small testicles. The large testicles displayed all germ cells at the steps of spermatogenesis. The small testicles presented no sperm as the animals housed in SP for 8 weeks. Thus these results indicate that acetaminophen invokes the antigonadal effects and accelerates the regressing process of the testicles in the animals compared to the animals exposed to SP.

Role of macrophage AHR/TLR4/STAT3 signaling axis in the colitis induced by non-canonical AHR ligand aflatoxin B1

Here we report that macrophage AHR/TLR/STAT signaling axis is implicated in the colon colitis induced by non-canonical AHR ligand aflatoxin B1 (AFB1). In BALB/c mice gavaged with 5, 25 and 50 µg/kg body weight/day AFB1, we observed severe colitis featured by over-recruitment of myeloid lineage immune cells such as monocytes/macrophage in colon lamina propria. Stressed and damaged colon epithelial cells were observed in low-dose group, while twisted and shortened intestinal crypts being found in middle dose group. Severe tissue damage was induced in the high-dose group. Dose-dependent increases of ROS, NO, and decrease of mitochondrial ROS-suppressor STAT3 were observed in the exposure groups. Further investigation in AFB1-treated human macrophage model found: (1) functional adaptations such as elevation of TNF-alpha and IL-6 secretion, stimulation of phagocytosis, elevation of LTE4 level; (2) overall inflammatory status confirmed by RNA-sequence analysis, in line with up-regulation of immune functional proteins such as ICAM-1, IDO-1, NF-kB-p65, NLRP3, COX-2 and iNOS; (3) mRNA disruption of mitochondrial oxidative phosphorylation complex I units and STATs; (4) perturbation of AHR/TLR/STAT3 signaling axis, including elevated AHR, TLR2, TLR4, and decreased STAT3, p-STAT3 Ser727. Mechanism investigation revealed regulatory links of ligand-dependent AHR/TLR4/STAT3. AHR-TLR4 together regulate MyD88, and STAT3 may be directly regulated by MyD88 (TLR4 downstream molecule) upon AHR/TLR4 binding with ligands. Solely protein level changes of AHR/TLR4 cannot regulate STAT3. Our study suggests that macrophage AHR/TLR4/STAT3 is involved with the colitis induced by sub-acute exposure to AFB1. Future follow-up study will focus on the intervention of the colitis using AHR-anti-inflammatory ligands.

A Phase 2b, Randomised, Double-blind, Placebo-controlled, Parallel-arm, Multicenter Study Evaluating the Safety and Efficacy of Tesnatilimab in Patients with Moderately to Severely Active Crohn's Disease.

Tesnatilimab, a monoclonal antibody targeting NKG2D, was evaluated in Crohn's disease [CD] patients who had failed or were intolerant to biologic or conventional therapy. TRIDENT was a phase 2b, two-part, randomised, double-blind, placebo-controlled, parallel-arm, multicenter study. In Part 1 [proof of concept], 145 patients who were biologic intolerant or refractory [Bio-IR] or had not failed biologic therapy [Bio-NF] were randomised in a 1:1 ratio to placebo subcutaneously [SC] or tesnatilimab 400 mg SC. In Part 2 [dose ranging], 243 Bio-IR and Bio-NF patients were randomised in a 1:1:1:1:1 ratio to placebo, tesnatilimab [50 mg, 150 mg, 400 mg], or intravenous infusion of ustekinumab ~6 mg/kg at Week 0 and 90 mg SC at Weeks 8 and 16. The primary endpoint was mean change from baseline in Crohn's Disease Activity Index [CDAI] at Week 8 [Part 1] and Week 12 [Part 2]. Clinical and endoscopic remission/response were evaluated. Efficacy analyses were also assessed by NKG2D and MICB single nucleotide polymorphism [SNP] status [SNP-positive means positive in at least one of two SNPs]. Safety events were summarised. In Part 1, mean change from baseline in CDAI score was significantly greater with tesnatilimab vs placebo at Week 8 [-103.6 vs -60.0; p < 0.01]. In Part 2, no dose-response signal was detected. Mean changes from baseline in CDAI at Week 12 were -93.2, -72.2, and -84.3 for low, middle, and high doses of tesnatilimab, respectively, vs -59.2 for placebo and -148.8 for ustekinumab. Similar reductions from baseline in CDAI score were observed in patients receiving tesnatilimab, regardless of SNP status. Clinical remission rates were greater with tesnatilimab than placebo in Parts 1 and 2, whereas endoscopic response rates were greater with tesnatilimab only in Part 1. No unexpected safety events occurred. Tesnatilimab was well tolerated. The efficacy of tesnatilimab in patients with CD was significant for the primary endpoint in Part 1; however, no dose-response signal was detected for the primary endpoint in Part 2. Based on these inconsistent findings, tesnatilimab was not considered an effective treatment for patients with CD and no further development is planned. NCT02877134.

Polysaccharides from small black soybean alleviating type 2 diabetes via modulation of gut microbiota and serum metabolism

Type 2 diabetes (T2DM) is a complex metabolic disorder associated with chronic high blood sugar and insulin resistance. Small black soybean, as a medicinal soybean, is used for tonifying kidney and spleen in traditional Chinese medicine. We performed the antidiabtic effect and mechanism of crude polysaccharides extracted from small black soybean (SB) by gut microbiota and metabolic modulation in STZ-induced diabetic rats. It is found that SB improved symptoms of hyperglycemia and hyperlipidemia and regulated linoleic acid metabolism. In addition, the relative abundance of Ruminococcus, Oscillospira, Dorea and Mucispirillum were modified by the middle dose of SB and these bacterial genera showed strong association with 6 vital metabolites and 10 risk indicators of T2DM. Additionally, it is further showed that 400 mg/kg pure polysaccharides exhibited more favorable antidiabetic activity than crude polysaccharides, which suggest that the antidiabetic effect of small black bean polysaccharides attributes to the pure polysaccharides.

The Effects of Ethanolic Extract of Premature Carica papaya (Pawpaw) Fruits on the Reproductive System of Female Wistar Rats

In recent times, different plant extracts have been reported to exert fertility enhancing effects. Carica papaya fruit is known for its use in preparation of herbal remedies. The aim of this study is to investigate the effects of Carica papaya on the reproductive system of female Wistar rats. Sixteen (16) female Wistar rats with regular oestrous cycle were used for this study. Animals were grouped into four (4) groups (control, low dose, middle dose, and high dose) of 4 animals each. Carica papaya fruit extract was administered for 14 days while monitoring the oestrous cycle. Twenty hours after the last administration, animals were sacrificed, and blood samples taken. Blood samples were collected for hormonal assay. Data obtained was expressed as Mean ± standard deviation, values were considered statistically significant when P ≤ 0.05. Phytochemical analysis of premature Carica papaya fruit revealed the presence of alkaloids, terpenes, sterols and terpenoids were heavily present. Estrous cycle was not altered. Serum FSH and LH increased. The extract is likely to possess fertility enhancing properties.

Neuroprotective effect of Dl-3-n-butylphthalide against ischemia-reperfusion injury is mediated by ferroptosis regulation via the SLC7A11/GSH/GPX4 pathway and the attenuation of blood-brain barrier disruption.

Stroke is one of the most severe diseases worldwide, resulting in physical and mental problems. Dl-3-n-butylphthalide, a compound derived from celery seed, has been approved for treating ischemic stroke in China. No study has evaluated how Dl-3-n-butylphthalide affects the ferroptosis SLC7A11/GSH/GPX4 signal pathway and blood-brain barrier (BBB) PDGFRβ/PI3K/Akt signal pathways in the rat middle cerebral artery occlusion/reperfusion (MCAO/R) model of ischemic stroke. Sprague-Dawley rats were used to develop the MCAO/R model. Our study used three incremental doses (10, 20, and 30) of Dl-3-n-butylphthalide injected intraperitoneally 24 h after MCAO/R surgery. The neuroprotective effect and success of the model were evaluated using the neurofunction score, brain water content determination, and triphenyl-tetrazolium chloride-determined infarction area changes. Pathological changes in the brain tissue and the degree of apoptosis were examined by hematoxylin and eosin, Nissl, and terminal deoxynucleotidyl transferase dUTP nick end labeling staining. In addition, pathway proteins and RNA expression levels were studied to verify the effects of Dl-3-n-butyphthalide on both pathways. At the same time, commercial kits were used to detect glutathione, reactive oxygen species, and malondialdehyde, to detect oxidative stress in brain tissues. The middle dose of Dl-3-n-butylphthalide not only improved MCAO-induced brain dysfunction and alleviated pathological damage, brain inflammatory response, oxidative stress, and apoptosis but also protected against ferroptosis and reduced BBB damage. These changes resulted in improved neurological function in the cerebral cortex. We speculate that Dl-3-n-butylphthalide has a neuroprotective effect on focal cerebral ischemia/reperfusion, which may be mediated through ferroptosis-dependent SLC7A11/GSH/GPX4 signal pathway and PDGFRβ/PI3/Akt signal pathway.