Introduction: Leydig cells (LCs) are the principal source of testosterone in males, underpinning male reproductive health and androgendependent physiology. Precise isolation of viable human LCs is essential for mechanistic steroidogenesis research and cell-based therapeutic development. Existing protocols rely on density-gradient centrifugation, which is technically demanding and often compromises yield and viability. Here, we describe a simplified density-gradient-free approach using differential adhesion to enrich functional human LCs from testicular tissue. Methodology: Human testicular fragments (~5 mm³) were minced and enzymatically digested with collagenase IV (2 mg/mL) at 37°C for 20 min under gentle agitation (100 RPM). The suspension was filtered (45 µm) and plated onto poly-L-lysine-coated T25 flasks. After 24 hours, non-adherent cells were removed by PBS washing. Viability exceeded 90% by trypan blue exclusion. Cells were maintained in DMEM/F12 with 10% FBS, 1% Antibiotic-Antimycotic, and 10 ng/mL luteinizing hormone (LH) to preserve the mature LC phenotype. Characterization employed immunofluorescence and flow cytometry using antibodies against SF-1, StAR, LHCGR, PDGFRA, and TEM-1. Testosterone secretion was quantified by ELISA under basal conditions and LH-stimulated conditions (10 ng/mL). Results: The protocol yielded ~1.25 × 10⁶ LCs per gram of tissue with >80% purity. Adherent HLCs displayed dense cytoplasmic lipid granules and intercellular networks consistent with active steroidogenesis. Flow cytometry confirmed 84.7% StAR⁺ cells, indicative of a robust steroidogenic population. A distinct progenitor subpopulation (PDGFRα⁺/TEM-1⁺) comprising ~28% of primary cultures, suggests retention of regenerative capacity. Basal testosterone secretion averaged 8.88 ng/mL per 24 hours, confirming preserved functional activity post-isolation. Conclusion This differential-adhesion protocol efficiently isolates functional human LCs without density-gradient media. Multimodal validation integrating immunofluorescence, flow cytometry, and ELISA confirms both phenotypic identity and steroidogenic competence, providing a reproducible and accessible platform for LC research and translational applications in male hypogonadism and androgen replacement.
Leydig Cells

Introduction: Capivasertib is a selective oral AKT inhibitor approved for hormone receptor-positive, HER2-negative advanced breast cancer with AKT pathway mutations. As AKT is integral to insulin signaling, its inhibition predisposes patients to hyperglycemia. We report a case of severe capivasertib-induced hyperglycemia with profound insulin resistance in a non-diabetic patient. Case: A 56-year-old female with metastatic breast cancer (ER/ PR-positive, HER2-negative, AKT1- and ESR1-mutated, TMB-high) presented with severe hyperglycemia following capivasertib initiation. She had no prior diabetes history, with a baseline hemoglobin A1c of 5.6%. Her first cycle (200 mg twice daily, 7 April 2025) was uncomplicated metabolically. The second cycle was escalated to 400 mg twice daily on 21 April 2025. By Day 3, blood glucose rose significantly. Premixed insulin 20 units failed to achieve glycemic control, necessitating intravenous insulin infusion. Despite escalation to 30 units per hour, euglycemia could not be achieved. Notably, there was no biochemical evidence of diabetic ketoacidosis or hyperosmolar hyperglycemic state, and the patient remained hemodynamically stable. No corticosteroids or other contributing medications were administered. Blood glucose normalized spontaneously approximately 36 hours after the last capivasertib dose, and all insulin was weaned and discontinued by 26 April 2025. She remained euglycemic without antidiabetic therapy thereafter. The patient and family opted for palliative care, and she passed away on 6 May 2025. Conclusion Capivasertib can precipitate severe, insulin-resistant hyperglycemia even in non-diabetic patients, without progression to overt hyperglycemic crisis. Spontaneous resolution upon drug cessation supports a direct drugmediated mechanism via AKT-disrupted insulin signaling. Clinicians should monitor glucose closely during therapy and consider drug cessation in refractory cases. Further studies are warranted to guide optimal glycemic management.
capivasertib ; Hyperglycemia ; Insulins

Introduction: Fulvestrant, a selective estrogen receptor degrader widely used in hormone receptor-positive breast cancer, can cause significant analytical interference with estradiol immunoassays due to structural similarity with 17β-estradiol. This interference may result in falsely elevated estradiol measurements, potentially leading to diagnostic confusion and inappropriate clinical interventions. We report a case demonstrating this clinically significant analytical interference and emphasize the importance of liquid chromatography-tandem mass spectrometry (LC-MS/MS) in hormone monitoring for patients receiving fulvestrant therapy. Case: A 31-year-old female with metastatic hormone receptorpositive breast cancer receiving combination therapy (fulvestrant, letrozole, ribociclib, and goserelin) presented with inappropriately elevated serum estradiol levels of 566 pmol/L (early follicular phase 200–500 pmol/L), measured by chemiluminescence immunoassay. Despite effective gonadotrophin suppression evidenced by low luteinizing hormone (0.2 IU /L) and follicle-stimulating hormone (4.1 IU/L) levels and clinical amenorrhea, the discordant estradiol elevation raised suspicion of assay interference. Confirmatory testing with LC-MS/MS revealed an estradiol concentration of <36 pmol/L, consistent with expected hormonal suppression and confirming immunoassay cross-reactivity with fulvestrant. The case demonstrates significant fulvestrant interference with estradiol immunoassays, resulting in a greater than 15- fold overestimation compared to LC-MS/MS measurement. The clinical presentation was consistent with effective endocrine therapy, supported by suppressed gonadotropin levels, clinical amenorrhea, and stable disease on imaging. LC-MS/MS provided accurate estradiol measurement, avoiding potential diagnostic confusion and unnecessary clinical interventions. Conclusion Clinicians should be aware of potential fulvestrant interference with estradiol immunoassays when interpreting hormone levels in breast cancer patients. Discordant laboratory findings, particularly elevated estradiol despite clinical evidence of effective hormonal suppression, should prompt consideration of LC-MS/MS confirmation. This case underscores the clinical importance of analytical method selection in hormone monitoring and highlights the need for improved laboratory-clinical communication to optimize patient care in oncology practice.
Fulvestrant ; Immunoassay

Epigallocatechin-3-gallate (EGCG), a bioactive polyphenol abundant in green tea, has garnered significant attention for its diverse therapeutic applications, ranging from antioxidant and anti-inflammatory effects to potential anticancer properties. Despite its immense promise, the practical utilization of EGCG in therapeutic settings as a medication has been hampered by inherent limitations of this drug, including poor bioavailability, instability, and rapid degradation. This review comprehensively explores the current challenges associated with the application of EGCG and evaluates the potential of nanoparticle-based formulations in addressing these limitations. Nanoparticles, with their unique physicochemical properties, offer a platform for the enhanced stability, bioavailability, and targeted delivery of EGCG. Various nanoparticle strategies, including polymeric nanoparticle, micelle, lipid-based nanocarrier, metal nanoparticle, and silica nanoparticle, are currently employed to enhance EGCG stability and pharmacological activity. This review concludes that the particle sizes of most of these formulated nanocarriers fall within 300 nm and their encapsulation efficiency ranges from 51% to 97%. Notably, the pharmacological activities of EGCG-loaded nanoparticles, such as antioxidative, anti-inflammatory, anticancer, and antimicrobial effects, are significantly enhanced compared to those of free EGCG. By critically analyzing the existing literature and highlighting recent advancements, this article provides valuable insights into the promising prospects of nanoparticle-mediated EGCG formulations, paving the way for the development of more effective and clinically viable therapeutic strategies.
Animals ; Humans ; Anti-Inflammatory Agents/administration & dosage* ; Antineoplastic Agents/administration & dosage* ; Antioxidants/administration & dosage* ; Biological Availability ; Catechin/analogs & derivatives* ; Micelles ; Particle Size ; Nanoparticle Drug Delivery System/chemistry*

Epigallocatechin-3-gallate (EGCG), a prominent plant-based catechin predominantly derived from Camellia sinensis and widely available on the market as a health supplement, has garnered significant attention for its potential therapeutic benefits, particularly in the context of type 2 diabetes mellitus (T2DM). This review explores the multifaceted role of EGCG in addressing the "ominous octet"-the 8 core pathophysiological defects associated with T2DM. The literature search was carried out using key terms "EGCG" OR "epigallocatechin-3-gallate" OR "epigallocatechin gallate" AND "diabetes" OR "insulin resistance" OR "hyperglycemia" in the PubMed and Scopus databases. The search was constrained to articles published between January 2018 and April 2024, focusing on the document type. Full-text articles published in English and relevant to EGCG that featured a single active ingredient, included clearly explained diabetes relief mechanism, and included ominous octet aspects were included in the final review. The outcomes of the included studies were reviewed and categorized based on 8 core pathophysiological defects, collectively referred to as the ominous octet in T2DM. This review concludes that EGCG is a potent hypoglycemic agent that has beneficial effects against the ominous octet in addition to its pharmacological activities in modulating gut microbiota dysbiosis, carbohydrate digestion and metabolism, glucose transporter-mediated intestinal glucose-uptake, endothelial dysfunction, and renal damage that are significantly associated with pathogenesis of T2DM. This extensive scientific evidence suggests that EGCG may offer a novel approach to traditional antidiabetic therapies, potentially improving glycemic control and mitigating complications associated with T2DM. The inhibitory effects of EGCG on sodium-glucose transport proteins and their role in reducing renal glucose reabsorption remain unexplored, highlighting a significant research gap. Future research should also aim to broaden the scope by investigating the "egregious eleven," which comprise a more comprehensive range of diabetic pathophysiological features. This review underscores the therapeutic promise of EGCG for managing T2DM and encourages ongoing research to fully elucidate its clinical applications. Please cite this article as: Wong CN, Lim YM, Liew KB, Chew YL, Chua AL, Lee SK. A review on mechanistic actions of epigallocatechin-3-gallate in targeting the ominous octet of type 2 diabetes mellitus. J Integr Med. 2025; 23(4): 344-356.
Diabetes Mellitus, Type 2/physiopathology* ; Humans ; Catechin/therapeutic use* ; Hypoglycemic Agents/therapeutic use* ; Animals ; Insulin Resistance

OBJECTIVE

Previous studies have indicated that clinical hypogonadism is common among males with type 2 diabetes mellitus (T2DM). However, the reported prevalence varies due to the diverse diagnostic criteria used in these studies. This study aims to determine the prevalence of clinical hypogonadism among Malaysian T2DM males and their associated factors.

A total of 360 participants who fulfilled the inclusion criteria were included in this study. Their socio-demographic and clinical parameters were documented and a total testosterone level was sampled from a morning fasting serum. Patients with serum total testosterone of 8-12 nmol/L had their serum total testosterone repeated and their symptoms assessed with the Aging Male Symptoms (AMS) scale. Clinical hypogonadism was diagnosed with total testosterone 26.

The prevalence of clinical hypogonadism among Malaysian T2DM males was 17.5% (n = 63), with 55.6% of them having hypogonadotropic hypogonadism. There is a significant association between clinical hypogonadism with waist circumference > 94 cm (p < 0.001), obesity (p < 0.001), hypertension (p = 0.010), coronary artery disease (p = 0.014) and peripheral artery disease (p = 0.022). There is a significant difference in the weight (p = 0.001), BMI (p < 0.001), waist circumference P < 0.001), serum HDL-C levels (p < 0.001), serum triglycerides levels (p = 0.001) and serum TyG index (p < 0.001). Diabetic males with increasing age (adjusted OR = 1.070, 95% CI 1.004-1.146, p = 0.038), presence of coronary artery diseases (adjusted OR = 2.08, 95% CI 1.220-10.219, p = 0.020) and low total testosterone (adjusted OR = 2.451, 95% CI 1.908-3.155, p < 0.001) are at higher risk of developing clinical hypogonadism.

This study is the first in the Asian region to use stricter criteria for diagnosing hypogonadism. Despite these stringent criteria, the prevalence of hypogonadism remains significantly high among Malaysian T2DM males. It is particularly common in diabetic males over 35 years old with coronary artery disease, regardless of A1c control and the duration of diabetes.

Rare Diseases-Malaysia.

Introduction: Time away from training and competition from sport injuries may lead to detrimental effects on muscle strength and endurance. The cross-education effect plays an important role in preserving strength during recovery and rehabilitation; however, the effects have been found to be inconclusive. In addition, the distinct impacts of eccentric and concentric strengthening exercises need to be explored. The objective of this study was to compare the cross-education effects of eccentric and concentric wrist flexor strengthening exercises on hand grip strength among recreational athletes. Methods: A total of 39 recreational athletes aged between 18-25 years old were randomly assigned into two groups of wrist flexor strengthening exercise: eccentric (ECC) and concentric (CON) training groups. The training period for the study was 4 weeks with 3 sessions of strengthening exercises per week. Handgrip strength was measured bilaterally, before and after the intervention using the JAMAR handgrip dynamometer. Results: Significant increases in hand grip strength were observed for the ECC and CON groups in both the trained and untrained hands following 4 weeks of training. However, the handgrip strength between the two groups in the untrained hand was not significantly different (p=0.64). The strength gain in the untrained hand was 9.8% and 10.8%, for ECC and CON groups, respectively. Conclusion: The cross-education effect was significant in improving strength in the untrained hand. There was no difference in strength gains between concentric and eccentric strength training.

Oral route drug delivery system is still considered as the most convenient and patient friendly drug delivery route. Over the decades, many research has been performed to improve the functionality oral dosage form. Orally disintegrating film (ODF) is a newer oral drug delivery system, which is in the form of a thin film that will disintegrate in the oral cavity within a matter of seconds. The aim of this review paper is to recap ODF, its benefits, formulation contents and manufacturing method. With more research and development work has been conducted on ODF, the dosage form is expected to be manufactured and scaled up to be commercializable products to be sold in the market.

Breast Conserving Surgery Post Neo-Adjuvant Chemotherapy for Locally Advanced Breast Carcinoma in a Developing Country: Is It Safe? Breast conserving surgery (BCS) is the standard treatment for early breast cancer and has similar survival with mastectomy. The role of BCS in locally advanced breast cancer (LABC), post neoadjuvant chemotherapy (NACT), is controversial. Surgeons, especially in developing countries, fear higher margin involvement and local recurrence (LR) in BCS. The aim of this study was to determine the LR in BCS compared to mastectomy in LABC post NACT and to ascertain the percentage of involved surgical margins following both methods of surgery. This was a retrospective study of breast cancer patients seen in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). All patients had NACT followed by either mastectomy or BCS. The patients with Her2 enriched carcinomas did not have access to targeted therapy (Trastuzumab or Pertuzumab). The patients with ill-defined tumours underwent ultrasound assessment in the surgical clinics. All had post-operative radiotherapy. All with ER or PR positive cancers were given Tamoxifen for a minimum of 5 years. This study demonstrated that BCS post NACT was safe to be performed in selected patients with LABC in a developing country.