The global surge in obesity, metabolic syndrome, and type 2 diabetes has driven a corresponding rise in non-alcoholic steatohepatitis (NASH), an advanced form of non-alcoholic fatty liver disease (NAFLD). Despite advancements in understanding its complex pathophysiology, NASH treatment options are limited. Current interventions primarily involve lifestyle changes, such as weight loss and exercise, and pharmacological agents which often present adverse effects, and no medication has been approved specifically for NASH. This highlights the urgent need for alternative therapeutic strategies. Tualang Honey (TH), a natural Malaysian product rich in antioxidants such as flavonoids and phenolic acids, represents a potential option for better NASH management. Previous studies have demonstrated TH's efficacy in improving liver enzymes and lipid profiles, particularly in animal models. However, the precise molecular mechanisms underlying its hepatoprotective properties remain poorly understood. This study investigated the molecular mechanisms underlying the attenuation of NASH through TH supplementation in a high-cholesterol diet (HCD)-induced NASH animal model. Twenty-five male Sprague Dawley rats were divided into five groups of 5 rats each: a normal control group (Group 1) fed with normal diet for 20 weeks and four groups fed a 12% HCD for 16 weeks to induce NASH, followed by four weeks of 12% HCD without TH (Group 2), 12% HCD and TH supplementation at doses of 1.2 g/kg/day (Group 3), moderate dose of 2.4 g/kg/day (Group 4), and high dose of 3.0 g/kg/day (Group 5). Histological examinations, biochemical assessment, immunohistochemical analyses, and gene expression profiling were conducted, followed by pathway analysis and validation of targeted gene expression. Statistical analysis was performed using one-way analysis of variance (ANOVA), followed by Tukey's least significant difference (LSD) post hoc test to identify group differences. The 12% HCD induced mild NASH, as evidenced by increased relative liver weight, presence of steatosis and inflammation in liver tissue samples, hypercholesterolaemia, hypertriglyceridaemia, as well as increased ?-SMA expression and TUNEL assay positive staining in the liver sections of Group 2. TH supplementation at 2.4 g/kg/day dose improved liver gross morphology, significantly reduced total triglycerides, activated hepatic stellate cells and apoptotic liver cells. Microarray analysis revealed ten most significant upregulated and 10 most downregulated genes involved in oxidative stress, immune response, lipid metabolism, and cellular injury repair in the TH-treated animals. Further pathway analysis demonstrated three key mechanisms underpinning TH's hepatoprotective effects: (1) lipid and cholesterol metabolism (2) antioxidant and anti-inflammatory pathways, and (3) glucose and energy metabolism. Notably, the significant upregulation of the aldehyde dehydrogenase (Aldh) gene at 2.4 g/kg/day compared to the HCD group indicates enhanced aldehyde detoxification processes with TH supplementation. Meanwhile, a significantly reduced suppression at 2.4g/kg/day of the interferon regulatory factor 2 (Irf2) gene aligns with an attenuated inflammatory response. Conversely, high-dose TH (3.0 g/kg/day) exhibited paradoxical effects, as the histology, biochemical profiles and immunohistochemical analysis demonstrate findings resembling NASH pathology. In conclusion, this study demonstrates that the hepatoprotective effects of TH in a 12% HCD-induced NASH animal model were most pronounced at the moderate dose of 2.4 g/kg/day. Lipid and cholesterol metabolism, antioxidant and anti-inflammatory mechanisms, and glucose and energy metabolism pathways were identified as the primary molecular pathways supporting these effects, highlighting the therapeutic potential of TH at optimal dosage.

Calcium phosphate is an ideal bone substitute material that is widely used for bone repair due to its excellent biological properties including biocompatibility and osteoconductivity. In order to improve the properties of calcium phosphate materials for clinical use, a new injectable self-hardened synthetic bone cement (Osteopaste) was developed. Osteopaste consists of tetra-calcium phosphate (TTCP) and tricalcium phosphate (TCP) powder. It was intended for the treatment of bone fracture or reconstruction of bone defects. The objective of this study was to compare bone formation between Osteopaste and commercialized synthetic bone grafts; JectOS (calcium phosphate) and MIIG-X3 (calcium sulphate) at three different assessment periods. The first phase of the study was to establish the critical size defect in New Zealand White rabbit model. The second phase involved the implantation of Osteopaste, JectOs and MIIG-X3 in critical size defects. Thirty-nine New Zealand White rabbits were divided into four groups (Osteopaste, JectOs, MIIG-X3 and sham). Each group was further divided into three subgroups according to the assessment period either at 6, 12 or 24 weeks. Each subgroup consisted of four rabbits except the sham group which consisted of only one rabbit. A critical size defect of approximately 4.5 mm (width) X 9.0 mm (length) was created at the proximal tibial metaphysis of rabbit's right leg and then implanted with either Osteopaste, JectOs or MIIG-X3. At each assessment period, plain radiograph and computed tomography (CT) scan were performed before the animals were sacrificed for undecalcified histology, histomorphometry and scanning electron microscopy assessments. Using the histomorphometric data, the mean percentage of new bone areas and the length of unbridged defects were compared between groups. In this study, a simple and safe method for performing critical size defect at proximal tibial metaphysis was established. The Osteopaste group exhibited radiographic density in between JectOS and MIIG-X3. The critical size defect in Osteopaste group was bridged by new bone at 12 weeks. In MIIG-X3 group, the defect was bridged at 24 weeks whereas in JectOS group, the defect was not bridged at all assessment periods. New bone area was the largest in MIIG-X3 group followed by Osteopaste and JectOS groups. Osteopaste had formed direct bonding with host bone without intervening soft tissue compared to JectOS and MilG-X3. There were significant differences in new bone area percentages between Osteopaste, JectOs and MIIG-X3 at 6, 12 and 24 weeks post-surgery (P<0.0001 ). In conclusion, the performance of Osteopaste to promote new bone formation is in between JectOS and MIIG-X3.

Parkinson’s disease (PD) is the most common motor disorder of the brain, with its global prevalence increasing by 153% since 1990. It is a progressive neurodegenerative disorder characterized by dopaminergic neuronal loss in the substantia nigra, with mitochondrial dysfunction and neuroinflammation as key pathological contributors. High-mobility group box 1 (HMGB1), a nuclear protein that acts as a damage-associated molecular pattern (DAMP) upon extracellular release, is implicated in both processes; however, its role in PD pathogenesis and therapeutic targeting remains underexplored. This study aimed to (i) refine behavioural and intraperitoneal (i/p) injection protocols for adult zebrafish, (ii) validate an MPTP-induced zebrafish model of PD at behavioural and transcriptional levels, and (iii) elucidate the functional role of HMGB1 and the impact of its inhibition using glycyrrhizin. The research was conducted in three phases. Phase 1 systematically reviewed existing MPTP-zebrafish protocols and optimized key methodologies for reproducibility, particularly locomotor assessment and i/p injection procedure. Phase 2 validated the PD model: MPTP administration induced significant hypo-locomotion and altered expression of dopaminergic marker genes (th, slc6a3) and PD/mitophagy-associated genes (pink1, prkn), alongside dynamic regulation of hmgb1 gene. Phase 3 examined HMGB1 modulation: glycyrrhizin treatment attenuated locomotor deficits, reduced upregulation of inflammatory mediators (TLR4, NFκB, TNFα, IL-1β), and normalized mitochondrial stress markers, while mitigating blood-brain barrier disruption. Tissue morphological alterations in the posterior tuberculum and subpallium persisted, suggesting incomplete structural recovery. In silico analysis confirmed conserved glycyrrhizin binding to HMGB1 in zebrafish and humans, reinforcing translational relevance. Findings indicate that HMGB1 is a central mediator linking neuroinflammation and mitochondrial dysfunction in MPTP-induced parkinsonism. Its pharmacological inhibition alleviated functional and molecular impairments, highlighting HMGB1 as a promising biomarker and therapeutic target for PD. This work also contributes standardized zebrafish methodologies, enhancing model reproducibility for neurodegenerative research. Keywords: Parkinson’s disease, HMGB1, adult zebrafish model, MPTP, neuroinflammation, mitochondrial dysfunction