Biomedical Research and Therapy

Association of stress and anxiety with Alzheimer’s: post-COVID-19 pandemic global perspective

Muhammad Khan — 2024-09-30

Alzheimer's is a neurodegenerative disorder mainly caused by aging, diabetes, stress, depression, anxiety, hypertension, obesity, etc. This bibliometric study investigated the correlation of stress and anxiety with Alzheimer's Disease to understand recent research trends and post-COVID-19 pandemic trending topics. The Web of Science database was utilized. The analysis was performed using the R package and VOSviewer software. A total of 305 articles were characterized and analyzed. The density visualization indicates that Alzheimer's disease and oxidative stress are the most prominent terms, followed by memory, dementia, anxiety, and stress. The most productive countries were China, the United States of America, and Iran.

Mutational variants of KRAS gene versus Wild-type KRAS in the survival outcomes of Vietnamese colon cancer stage II-III

Hoang Minh Cuong — 2024-09-30

Background: Colon cancer is one of the most common cancers in Vietnam and globally. The KRAS gene (Kirsten rat sarcoma) is an oncogene showing a high mutation rate in colon cancer, affecting 30% to 40% of patients. The mutated KRAS gene, which keeps the MAPK signaling pathway permanently active, is considered a negative factor in the survival of colon cancer patients. Recently, several studies have been conducted to evaluate the specific prognoses related to distinct KRAS mutations, but the results were controversial. Mutations in different KRAS codons may impact colon cancer treatment models and prognoses. Therefore, the impacts of codon-specific KRAS mutations on survival require further clarification. This study aims to determine the associations between codon-specific KRAS mutations and survival in Vietnamese patients at stages II - III.

Methods: A descriptive design was applied and included 158 colon cancer patients at stages II-III at Bach Mai Hospital from January 2016 to August 2020. Testing for KRAS mutations was performed with formalin-fixed, paraffin-embedded tissue, and KRAS mutations were detected with the KRAS XL StripAssay (ViennaLab, Austria).

Results: Among 158 patients, 71 (44.9%) exhibited mutated KRAS genes. The most frequent mutated KRAS variant was p.Gly12Asp (G12D) at 35.2%; the proportion of KRAS G13D (p.Gly13Asp) was 15.5%. The 3-year DFS of the wild-type KRAS group was 60.3%, while that of the mutated KRAS Exon 2 group was 55.2%. The worse prognosis for DFS was observed in patients with KRAS codon 12 mutations (3-year DFS: 52.9%). Among them, patients with codon-specific KRAS mutations named p.Gly12Asp (G12D) and p.Gly12Val (G12V) experienced better prognoses (59.1% and 63.6%, respectively). However, the differences were not statistically significant (P > 0.05).

Conclusion: KRAS p.Gly12Asp (G12D) was the most common type among mutated KRAS genes. Codon-specific KRAS mutation was not a prognostic factor for DFS and OS in colon cancer stages II-III. The results support the current clinical practice that determining specific codon KRAS gene status is not conventional testing for resected colon cancer stages II-III.

What is the optimal timing and solution for the intravenous infusion of thawed cryopreserved mesenchymal stem cells?

Trang Thi-Hien Nguyen — 2024-09-30

Introduction: The viability of mesenchymal stem cells (MSCs) during intravenous infusion can significantly impact treatment efficacy, particularly with thawed cryopreserved MSCs. During infusion, these thawed MSCs are suspended in saline at room temperature, which lacks nutrients and buffering capacity, leading to a gradual decline in cell viability. This study aimed to investigate the survival of thawed MSCs during intravenous infusion and propose an optimal procedure to maximize cell viability.

Methods: Human MSCs derived from adipose tissues (ADSCs) and umbilical cords (UCMSCs) were utilized in this study. The MSCs were cryopreserved using a cryoprotectant-free medium (MSCCryosave OTS) at -196°C at a concentration of 10⁷ cells/mL. They were then naturally thawed at room temperature. The 15.10⁷ thawed MSCs in MSCCryosave OTS were suspended in 500 mL Lactated Ringer solution. The bottle was connected to an infusion tube, with the valve opened at a rate of 25 drops/min. Cell viability was assessed every 30 minutes until the bags were depleted. In another experiment, cell viability was evaluated using a phosphate-buffered saline (PBS) solution supplemented with 5% human serum albumin (the solution named CellCarrier). Cell viability was determined using a hemocytometer with trypan blue staining.

Results: The results indicated that the cell viability of thawed ADSCs and UCMSCs gradually decreased over time. For ADSCs, the percentage of viable cells decreased from 92.95 ± 1.33% post-thaw (at 0 h) to 80.41 ± 2.02% at 90 minutes of transfusion. For UCMSCs, the percentage decreased from 94.44 ± 1.69% post-thaw (at 0h) to 81.12 ± 2.26% at 90 minutes of transfusion. At the last drop of the bag, the percentage of viable ADSCs and UCMSCs was 48.22 ± 14.08% and 59.39 ± 14.54%, respectively, after approximately 385 minutes of transfusion. Notably, cell viability significantly increased when cells were infused with CellCarrier, remaining above 90% after 385 minutes (90.13 ± 0.24% for UCMSCs and 90.09 ± 0.44% for ADSCs).

Conclusion: This study suggests that using Lactated Ringer's solution results in a rapid decline in cell viability. Therefore, reducing the transfusion time is crucial to maintaining cell viability before administration to patients. We recommend keeping the transfusion time under 90 minutes when using Lactated Ringer's or alternatively utilizing CellCarrier solution for infusion.

The proliferation of MDA-MB-231 cells is repressed by mesenchymal stem cell-mediated macrophage activation conditioned medium through the inhibition of AKT1 and YKL-39 genes

Nur Ramziahrazanah Jumat — 2024-09-30

Background: Triple-negative breast cancer (TNBC) is characterized by a substantial presence of tumor-associated macrophages (TAMs) exhibiting an M2-like phenotype, which plays a crucial role in promoting tumor cell stemness and invasiveness. Mesenchymal stem cells (MSCs) have the ability to induce the transformation of naive macrophages (M0) into M1-like macrophages. This study delves into the interplay between MSCs and macrophages within the context of breast cancer (BC) progression using a TNBC cell line, as reprogramming of TAMs into M1-like macrophages may emerge as a promising therapeutic strategy for BC.

Methods: THP-1 cells were induced into M0 macrophages and co-cultured with UC-MSCs, subsequently analyzing CM for M1- and M2-type macrophage-related cytokines. Total RNA from co-cultured cells was used to assess IRF-4 and IRF- 5 mRNA gene expression via qRT-PCR. MDA-MB-231 cells were exposed to CM and co-cultured cells to evaluate cell viability through MTT assay over 24, 48, and 72 hours, with qRT-PCR used to examine breast cancer-related gene expression.

Results: The results indicate that co-culturing M0 macrophages with MSCs promotes M1-like macrophages, as evidenced by upregulated IRF- 5 and suppressed M2 macrophage-related genes. Treatment with CM from M0/MSCs co-culture significantly inhibits MDA-MB-231 cell proliferation at 72 hours, accompanied by reduced TNF-alpha levels. Notably, CM treatment downregulates AKT1 and YKL-39 genes in MDA-MB-231 cells, suggesting potential anti-cancer effects. Direct co-culture with M0/MSCs, however, shows no significant impact on TNBC cell growth.

Conclusion: This study highlights MSCs' ability to induce M0 macrophages to a M1-like phenotype and suggests that CM from M0/MSCs co-culture may contain anti-cancer factors targeting AKT1 and YKL-39 genes, underscoring the potential of MSC-mediated macrophage activation as a strategy to enhance BC treatment, especially in the context of TNBC.

Predicting lymph node metastasis in gastric adenocarcinoma: Role of tumor budding and immunohistochemical expression of E-cadherin

Hung Van Nguyen — 2024-09-30

Introduction: Predicting the node status of stomach carcinoma is very useful, as it affects treatment decisions. The objective of this study was to use the immunohistochemical expression of E-cadherin and tumor budding to investigate the nodal status of gastric cancer surgical specimens as a pilot study before application to gastric cancer biopsy samples.

Methods: Three hundred and eleven (311) gastric cancer surgical samples with lymph node dissection were retrospectively evaluated at Hanoi Medical University Hospital, Vietnam. The comparison of tumor budding and E-cadherin expression with lymph node status was investigated. Tumor budding was calculated on the microfield of 0.785 mm² according to the 2016 International Consensus Conference guidelines. The immunoexpression of the E-Cadherin protein was examined by immunohistochemistry.

Results: The overall lymph node metastasis rate was 55.6%. In multivariable logistic regression analyses, tumor budding (odds ratio [OR] = 12.73, 95% confidence interval [CI] = 4.980–32.53, p < 0.001), immunoexpression of E-cadherin (OR = 0.048, 95% CI = 0.019–0.121, p < 0.001), stage (OR = 2.329, 95% CI = 1.204–4.504, p = 0.012), and grade (OR = 2.032, 95% CI = 1.081–3.820, p = 0.028) were critical factors that can independently predict lymph node status.

Conclusions: Tumor budding and E-cadherin expression are independent factors and may be additional candidates in predicting node status in patients with primary gastric carcinoma.

Effects of Orychophragmus violaceus extract OVS-2 on the structure and diversity of intestinal flora in mice with radiation intestinal injury

Haixia Li — 2024-09-30

Introduction: Orchophragmus violaceus (OV) has a rich history of use in traditional Chinese medicine, spanning millennia. Its diverse physiological properties, such as anti-radiation, antibacterial, antitumor, and hepatoprotective effects, have garnered widespread recognition. This study aims to elucidate the potential of OV extracts to mitigate radiation-induced intestinal injury (RIII) and its impact on the intestinal microbiota.

Method: OVS-2, extracted and isolated from OV seeds, was administered to C57 mice following abdominal irradiation (IR) with ⁶⁰Co rays for RIII. Tissue samples from the small intestine and fecal matter were collected from three groups of mice three days post-administration for histological examination (HE staining) and 16S rRNA fecal intestinal flora sequencing. The composition of the intestinal flora was analyzed through 16S rRNA gene amplification.

Result: In vivo experiments demonstrated that OVS-2 supplementation improved the survival rate of mice and mitigated radiation-induced damage to intestinal villi and crypts to some extent. The relative abundance of Firmicutes, Bacillus, and Lactobacillus decreased in the experimental group compared to the model group, while that of Actinobacteria, Erysipelotrichaceae, and Dunaliella increased. LEfSe analysis revealed an increased relative abundance of Faecalibacterium, Dunaliella, Erysipelotrichales, and other bacteria, indicating their potential importance in the mouse intestinal microbiota. Dysbiosis was evident in the intestinal flora of mice in the IR group.

Conclusion: Oral administration of OVS-2 can help re-establish the gut microbiota composition and slow down the development of RIII in mice, suggesting that OVS-2 can regulate RIII and disturbance of intestinal flora in mice.

Development of a recombinant construct as an immunogenic target for leptospirosis using LipL41 and molecular adjuvants

Govindan Pothiaraj — 2024-09-30

Introduction: Leptospirosis, an infectious disease that can spread from animals to humans, requires the development of a safe and effective vaccine. The immunogenic characteristics of LipL41, a conserved outer membrane protein of Leptospira, have been identified as a promising vaccine candidate. In this study, a recombinant DNA construct, pTR-EGFP-LipL41, incorporating the LipL41 gene and hGMCSF adjuvant in the pTR600 vector with a cytomegalovirus (CMV) promoter, was designed and evaluated.

Methods: The Chinese hamster ovary (CHO) cell line was transfected with pTR-EGFP-LipL41 and pTR-EGFP-hGMCSF-LipL41 using Lipofectamine 2000, and fluorescence microscopy analyzed their expression.

Results: The expression analysis demonstrated successful expression of pTR-EGFP-LipL41 and pTR-EGFP-hGMCSF-LipL41 in CHO cells. In vitro analysis of cell lines further assessed the expression of chemokines and cytokines, and molecular docking analyses were conducted to investigate interactions between various adjuvants (hGMCSF, hIgGFc, and hC3d) and LipL41. Docking studies uncovered key interactions between LipL41 and other adjuvants. The constructed recombinant DNA and molecular adjuvants exhibited a robust immunogenic response.

Conclusion: Further evaluation in suitable animal models may establish its effectiveness as a productive and safe immunogenic molecule against leptospiral infection.

MSCs, but not mesenchymal stem cells

Phuc Van Pham — 2024-09-30

Mesenchymal stem cells (MSCs) are prevalent within the human body and can be detected and isolated from nearly all tissues. The acronym MSCs is widely recognized as referring to adult stem cells with multipotent capabilities. Despite over 40 years of applications in research and clinical settings, the definition of MSCs as mesenchymal stem cells has faced scrutiny concerning their "stemness" and the underlying mechanisms of their therapeutic effects. In 2010, Dr. Arnold I. Caplan suggested redefining MSCs as "medicinal signaling cells" rather than mesenchymal stem cells. In this commentary, I concur with Dr. Caplan's view but further propose that MSCs be regarded as "master signaling cells." The primary therapeutic mechanism of MSCs is their signaling function. They respond to signals from immune cells to become activated and, in turn, act as signaling regulators for other cells. As master signaling cells, MSCs are multipotent stem cells present in almost all tissues, playing vital roles in regulating tissue homeostasis and facilitating tissue regeneration.

A fatal case of overwarfarinization leading to complete paraplegia with hematomyelia

Al Hafiz Ibrahim — 2024-09-30

Background: Warfarin is a commonly prescribed anticoagulant medication used for thromboprophylaxis in patients at an increased risk of thromboembolic events. However, warfarin administration can lead to hemorrhage due to its narrow therapeutic range. Hemorrhage may occur in unusual anatomical sites and can result in mortality.

Case Presentation: We report the case of an 84-year-old male patient diagnosed with non-valvular atrial fibrillation (AF) and hyperthyroidism, who presented with the sudden onset of lower back pain, bilateral lower limb weakness, and paraesthesia. He was diagnosed with an intramedullary epidural hematoma leading to spinal cord compression as a consequence of overwarfarinization. Following the administration of a new increased dose of oral warfarin, the patient experienced sudden onset back pain accompanied by complete paraplegia. After reversing the overwarfarinization to a therapeutic level, an emergency surgical procedure involving spinal decompression, clot removal, and posterior instrumentation was planned. However, the patient developed an onset of unstable tachyarrhythmia following the administration of anesthesia, leading to the discontinuation of the surgery. Due to substantial risks, the patient declined to undergo the surgical procedure. Despite a consistent neurological condition, he developed a disability and subsequently succumbed to death due to a respiratory infection within four weeks following the initial presentation of symptoms.

Conclusions: This case represents a rare instance of intramedullary epidural hematoma resulting in spinal cord compression due to overwarfarinization, highlighting the need for awareness and urgent management.

Neuropsychological manifestation as an initial presentation of pediatric chronic myeloid leukemia: A case report of diagnostic conundrum and review of literature

Anis Munirah Mohd Kori — 2024-09-30

Background: Chronic myeloid leukaemia is a form of myeloproliferative neoplasm characterised by the proliferation of granulocytes. This particular form of myeloproliferative neoplasm is rare, accounting for approximately 2% – 3% of recently diagnosed pediatric leukaemia. Neuropsychological manifestation is a rare initial sign in patients with chronic myeloid leukaemia.

Case presentation: A 10-year-old girl who presented with two weeks of abnormal behaviour. Her neurological assessment revealed no abnormality. Her serial full blood count revealed persistent leucocytosis, and her most recent full blood picture showed 2.3% blast cells. Following a comprehensive analysis of bone marrow smear and identification of the BCR-ABL fusion gene, it was concluded that the patient had chronic myeloid leukaemia in chronic-phase. Her neuropsychological symptoms improved after a few months of imatinib treatment, and she will be scheduled for a stem cell transplant if her human leukocyte antigen matches those of other siblings.

Conclusion: Neuropsychological manifestations in chronic myeloid leukaemia-chronic phase are uncommon in children and can be mistaken for more prevalent conditions. Physicians should maintain a vigilant stance due to the distinct management and prognosis of chronic myeloid leukaemia in comparison to the more prevalent condition in paediatric.