Annals of clinical and laboratory science最新文献

Objective: Non-alcoholic fatty liver disease (NAFLD) is a growing health concern with limited early treatment options. Puerarin, an active component derived from Pueraria lobata (kudzu), has been traditionally used in Chinese medicine for its potential therapeutic benefits. This study explores the effect of puerarin on the development of NAFLD and its mechanisms.

Methods: Sprague Dawley rats with NAFLD were induced with a high-fat diet (HFD). Control rats received a standard diet, while puerarin was administered by gavage (0.4 or 0.8 g/kg) daily from week 9. Body weights were recorded weekly. After 16 weeks, liver tissues and related indicators of rats were examined. Gut microbiota was assessed via 16S rRNA sequencing of fecal sample.

Results: Compared to HFD rats, those treated with puerarin showed significant reductions in body weight, liver weight, and liver index. Liver tissue levels of TG, TC, ALT, AST, and inflammatory factors were also significantly decreased. In addition, serum levels of TG, TC, and LDL-C were lower, while HDL-C level was higher in puerarin-treated groups. 16S rRNA sequencing revealed that gut microbiota composition and diversity in the puerarin groups resembled those of healthy rats, maintaining the level of Subdologranulum and decreased the level of Ruminococcus.

Conclusion: Puerarin alleviates HFD diet-induced NAFLD by reducing inflammatory cytokine production and modulating the gut microbiota. Puerarin is a potential therapeutic drug for NAFLD, and regulating the gut microbiota is an effective strategy for treating NAFLD.

Objective: To assess the performance of neutralization confirmatory testing for borderline hepatitis B surface antigen (HBsAg) results (signal-to-cutoff [S/CO] 1.0-10.0) in a low-prevalence population.

Method: We retrospectively analyzed 115 borderline-reactive HBsAg samples from asymptomatic adults with normal liver enzyme levels. Screening was performed using the Abbott Alinity i assay, followed by confirmatory testing with the Abbott HBsAg Confirmatory Reagent. A ≥50% signal reduction was required for confirmation.

Results: Among 115 borderline samples, 61 (53.0%) were confirmed by neutralization testing. Confirmation rates increased with higher S/CO values: 31.6% (1.0-2.0), 57.1% (2.1-5.0), and 75.0% (5.1-10.0). The median S/CO values were 1.5 (1.0-2.0), 3.4 (2.1-5.0), and 7.2 (5.1-10.0) for each subrange. Logistic regression showed a significant correlation between higher S/CO values and confirmation (odds ratio 1.42 per unit increase, p<0.001). Notably, most unconfirmed cases had S/CO values below 3.0, suggesting frequent false reactivity in low-positive samples.

Conclusion: Lower borderline HBsAg results often reflect nonspecific assay reactivity, emphasizing the need for confirmatory testing to prevent misdiagnosis. Higher S/CO values were strongly associated with true HBsAg positivity, supporting the use of neutralization testing in routine practice.

Objective: The peptide PNC-27 has been found to kill many different endodermal solid tissue and hematopoietic cancer cells but has no effect on normal cells. The mechanism involves binding to the HDM-2 protein, which is expressed in the membranes of cancer cells but not in normal (untransformed) cells. Our objectives in the current study are to determine 1) if PNC-27 is lethal to squamous cervical epithelial cancer cells but not to untransformed squamous cervical cells; 2) if membrane-bound HDM-2 is expressed uniquely in cervical cancer cells; and 3) whether HDM-2 is stable for detection in different types of preservative solutions.

Methods: We determined dose response curves for incubation of PNC-27 with the human squamous cervical cancer cell line HTB-35 (also called SiHa cells) and with the untransformed human squamous cervical cell line, PCS-480. Cell viability was determined using the MTT and LDH release assays. Finally, slot blots and flow cytometry were used to determine membrane expression of HDM-2 using a polyclonal anti-HDM-2 antibody.

Results: We found that PNC-27 is cytotoxic even at low doses (IC₅₀=12.4 μM) to the human HTB-35 cervical cancer squamous epithelial cell line but not to a counterpart normal human PCS-480 cell line. We found that HTB-35 cells express high levels of HDM-2 proteins in their membranes both in cell culture and in alcoholic preservative solutions but that the normal PCS-480 cells do not. Consistent with previous results, the data suggest that cervical cancer cells express HDM-2 in their membranes and that this is the target for PNC-27.

Conclusions: PNC-27 kills cervical squamous cancer but not normal cervical cells due to the unique expression of HDM-2 in the cervical squamous cell membranes. Thus, PNC-27 may be an effective drug against this cancer. Our results further suggest that the expression of membrane-bound HDM-2 on cervical cancer cells is stable both in cell culture media and in alcoholic preservative fluid.

Objective: Elevated serum immunoglobulin (Ig)E levels are associated with progression of human immunodeficiency virus (HIV) infection and altered T cell regulation. High serum IgE levels may be a clinical indicator of disease in HIV+ adults; however, few studies in children have been reported. The aim of this study sought to determine whether there exists a correlation between serum IgE levels, viral load, or CD4+ and CD8+ T cell in children living with perinatally acquired HIV (CPHIV) in Brooklyn, N.Y.

Methods: Pediatric patients (N=6, 67% female) diagnosed with HIV infection on anti-retroviral therapy. Age of entry (time 1) was 1-5 years old; follow up (time 2) was six years later. The primary analysis compared specific variables: total serum IgE (IU/mL), viral load (RNA copy/mL), CD3+, CD4+, and CD8+ T cells (%).

Results: Percentages of lymphocytes (CD3+, CD4+, CD8+) and serum IgE levels increased, but CD4/CD8 ratio and viral load decreased from time 1 to time 2. However, only changes in CD8+ T cells were significant (mean difference: -8.33(5.72), P=0.031) (Wilcoxon signed-rank test). At time 2, mean differences were not significant when subjects were stratified according to positive/negative serum IgE status or high/low viral load.

Conclusion: In CPHIV, CD8+ T cells significantly increased over time, confirming their importance as a marker for disease progression. However, the relevance of high serum IgE levels in CPHIV remains unclear. Understanding unique biomarkers in CPHIV is important for early life HIV cure strategies.

Objective: This study investigates the prevalence and classifications of breast tumors in the Asir region over the last five years, together with abnormal hematological parameters and coagulation profiles prior to cancer therapy.

Methods: This retrospective analysis, covering the period from 2018 to 2022, was conducted at Asir Central Hospital in Abha, Saudi Arabia. Data on demographics and tumor types were obtained from the medical records of 764 patients. Hematological parameters and coagulation profiles of 94 malignant breast cancer patients and control samples were compared using GraphPad Prism.

Results: The majority of cases were benign breast disease (61%, 473), followed by malignant tumors (38%, 292). The most common benign subtypes were fibroadenoma (53.2%, 252 patients), fibrocystic breast alterations (12.6%, 60 patients), and fibroadenosis (9.9%, 47 patients). Among malignant tumors, invasive ductal carcinoma (82.1%, 240 patients), ductal carcinoma in situ (7.1%, 21 patients), and invasive lobular carcinoma (3.7%, 11 patients) predominated. Malignancy patients had lower HB, RBC, MCHC, MCH, MCV, and HCT, and higher RDW. In addition, INR was significantly lower than the control group.

Conclusions: Over the five-year period ending in 2022, the incidence rate of malignant breast cancer increased in the Asir region. Patients with such cancers show significant abnormalities in hematological parameters and coagulation profiles prior to treatment.

Hemophagocytosis is the process of phagocytosis of erythrocytes or other hematopoietic precursors by histiocytes or macrophages. Increased histiocytic activity may be observed in infections, inflammation, bone marrow hyperplasia, ineffective hematopoiesis, malignancies, as well as in hemophagocytic lymphohistiocytosis (HLH). Here we present two challenging cases of acute myeloid leukemia (AML) with associated hemophagocytosis, one in which hemophagocytosis provided a diagnostic cue and another in which extensive HLH obscured the underlying AML. The first case highlights the characteristic morphologic (leukemic blasts with erythrophagocytosis) and immunophenotypic findings (high side scatter and bright HLA-DR expression) observed in AML with t(8;16). The second case highlights the importance of careful bone marrow examination to rule out an underlying malignancy in children presenting with HLH, as the diagnosis and treatment of primary malignancy is crucial in successful management. The second case also shows an uncommon presentation of AML with concomitant EBV-associated HLH. A high index of suspicion for malignancy based on clinical history and careful bone marrow examination led us to reach the definitive diagnosis in a timely manner and allowed for optimal patient management.

Objective: Esophageal squamous cell carcinoma (ESCA) is a prevalent malignant tumor with poor prognosis. Interleukin 2 receptor beta (IL2RB) has been implicated in various cancers; however, its role in ESCC remains unclear.

Methods: We analyzed IL2RB expression in clinical samples and cell lines. The impact of IL2RB on tumor progression was assessed using gain- and loss-of-function approaches, along with in vivo tumor models. In addition, we explored the effect of IL2RB on the immune microenvironment and its potential to modulate the JAK1/STAT5 pathway.

Results: IL2RB was found to be significantly upregulated in ESCC tissues compared to normal tissues. Functional studies revealed that IL2RB knockdown inhibited tumor cell proliferation, migration, and invasion, as well as epithelial-mesenchymal transition (EMT). IL2RB was shown to reshape the tumor immune microenvironment by inducing CD8+T cell depletion. Mechanistic investigations indicated that IL2RB activates the JAK1/STAT5 pathway, thereby promoting ESCC progression.

Conclusions: Our findings demonstrate that IL2RB plays a critical role in ESCC progression and immune evasion, suggesting its potential as a therapeutic target. Further studies are warranted to explore the clinical application of IL2RB targeting in ESCC treatment.

Objective: The purpose of this study was to explore the effects and potential mechanisms of miRNA-29c-3p regulating suppressors of cytokine signaling 3 (SOCS3) in Diabetic Retinopathy (DR) progression.

Methods: Human retinal microvascular endothelial cells (hRMECs) were exposed to 25 mM high glucose concentrations to establish a DR cell model and underwent transfection to down-regulate miRNA-29c-3p and SOCS3.

Results: High glucose induces upregulation of miRNA-29c-3p and downregulation of SOCS3 expression in hRMECs. Under high-glucose conditions, inhibition of miRNA-29c-3p significantly suppresses hRMEC migration, angiogenesis, and the release of pro-inflammatory cytokines. Notably, this inhibitory effect is partially reversed upon SOCS3 knockdown. Moreover, miRNA-29c-3p directly targets and regulates SOCS3 mRNA expression. Importantly, SOCS3 knockdown markedly activates the JAK2/STAT3 signaling pathway in hRMECs, which can be suppressed by reducing miRNA-29c-3p levels.

Conclusion: miRNA-29c-3p promotes DR progression by activating the JAK2/STAT3 pathway through SOCS3 regulation.