Annals of clinical and laboratory science最新文献

A 20-year-old pregnant woman at 12 week gestation with a history of thalassemia was admitted to the hospital with Hb 60g/L. She received two transfusions of 2 units of negative crossmatched washed red blood cells (RBCs) each, but shortly after she experienced a transfusion reaction. Symptoms included chest tightness, dyspnea, chills, and soy sauce colored urine. A post-transfusion specimen was sent to the blood type reference laboratory (BTRL) for investigation, which revealed the presence of anti-Le^a and anti-Le^b antibodies causing the immediate acute hemolytic transfusion reaction; interestingly, the patient's Le^a antibody was found to be IgM, while the Le^b antibody was both IgM and IgG. This combination of antibodies is rare and highlights the potential for clinically insignificant Lewis cold antibodies to cause serious reactions. It is important to not overlook these antibodies and to select antigen-negative units rather than relying solely on blood crossmatching. The use of polybrene in crossmatching blood tests may have limitations in the presence of Lewis antibodies, so alternative methods should be considered in difficult cases to ensure safe and effective transfusions. This case emphasizes the need for thorough testing and careful selection of blood products to reduce the risk of transfusion reactions and improve overall transfusion safety.

Objective: In approximately 80% of the patients with advanced non-small cell lung cancer (NSCLC), the only samples available are cytological material or a limited amount of tissue specimens. Therefore, we aimed to explore the effect of the Idylla™ system for rapid evaluation of epidermal growth factor receptor (EGFR) mutation status in formalin-fixed and paraffin-embedded (FFPE) samples of NSCLC patients.

Methods: A total of 221 archived FFPE NSCLC tissue specimens were included in this study. Idylla™ system and amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) were used to detect the EGFR mutation status. The Idylla™ system results and ARMS-PCR results were compared. When results were discordant, the Idylla™ system would be retested, and a third analysis method was performed to confirm test results when possible.

Results: Among the 220 valid results of the Idylla™ system, the Idylla™ identified EGFR mutations in 109 cases with an incidence of mutation of 49.55%. The Idylla™ assay results were in complete agreement with the results of the reference methods for 207 cases, yielding an overall accordance of 94.09% (207/220, 95% CI, 90.11%-96.82%). Consequently, the overall concordance with routine reference methods, including further analysis, was found to be 96.81% (213/220, 95% CI: 93.28%-98.60%), with a negative percentage agreement of 97.3% (108/111, 95% CI: 91.72%-99.30%) and a positive percentage agreement of 96.33% (105/109, 95% CI: 90.32%-98.81%).

Conclusion: The Idylla™ system is a rapid and effective method that enables sensitive and reliable detection of EGFR mutations in FFPE samples of NSCLC patients, with minimal molecular expertise or infrastructure.

Objective: MicroRNA-192-2 has been shown to have a role in the early diagnosis of hepatocellular carcinoma (HCC), but the relationship between microRNA-192-2 and hepatitis B virus (HBV) in HCC patients remains unclear.

Methods: Specimens were collected from 56 HCC patients diagnosed with HBV infection and 56 HCC patients without viral infection. HBV and miR-129-2 levels in HCC tissues, adjacent tissues, and cell lines were analyzed by RT‒PCR. miR-129-2 mimics were transfected to induce the overexpression of miR-129-2 and cell function was assessed by a wound healing test. Tumor formation experiments in nude mice were conducted to validate tumor proliferation.

Results: In HCC patients, miR-192-2 was significantly downregulated in tumor tissues compared to adjacent tissues. Notably, miR-192-2 level was even lower in HCC patients with HBV-infection than those without the viral infection. Moreover, there was a negative correlation between miR-192-2 and HBV levels. Regardless of HBV infection, patients with low miR-192-2 levels had poorer prognosis than those with high miR-192-2 levels. HBV infection suppressed miR-192-2 expression in HCC cell lines. Furthermore, overexpression of miR-192-2 significantly inhibited cell proliferation both in vitro and in vivo in the HBV-infected group.

Conclusion: Our study revealed that the expression of miR-192-2, a crucial tumor suppressor gene, was suppressed by the presence of HBV.

The case of a 49-year-old man with Hb Phnom Penh whose HbA1c levels varied depending on the high-performance liquid chromatography (HPLC) system is presented. After the results of a health checkup showed a high HbA1c level of 7.6% measured by HPLC method, another HbA1c test measured by immunoassay at a local clinic showed a level of 5.2%. Given the discrepancy in HbA1c levels between assay methods, he visited our clinic. His fasting plasma glucose level was 95 mg/dL, but an oral glucose tolerance test showed diabetes mellitus. The mean plasma glucose level was 117 mg/dL on intermittently scanned continuous glucose monitoring (isCGM), giving an estimated HbA1c level of 5.9%. Based on the results of isCGM, the present case was considered to be not diabetes mellitus. The same specimen was assayed for HbA1c by various methods (systems), including HPLC with Tosoh G9, HPLC with Arkray HA-8180 and HA-8180T, affinity assay, and enzymatic assay, giving HbA1c levels of 7.3%, 4.9%, 5.0%, 5.4%, and 5.3%, respectively. A globin gene analysis diagnosed Hb Phnom Penh (α1-117Phe-Ile-α1-118Thr). The present case had a unique hemoglobin variant, resulting in high HbA1c levels when measured with Tosoh's HPLC systems and low HbA1c levels with Arkray's HPLC systems.

Objective: To evaluate the effect of osteoprotegerin (OPG)/receptor activator of nuclear factor-[Formula: see text]B ligand (RANKL)/receptor activator of nuclear factor-[Formula: see text]B (RANK) nuclear factor on aortic valve calcification.

Methods: The aortic valve tissue was collected from 132 aortic stenosis (AS) patients who underwent valve replacement. The valve tissue was stained with hematoxylin eosin (HE) and alizarin red calcium salt deposition. At the same time, CD68, RUNX2, TRAP immunohistochemical staining and double staining were performed.

Results: ELISA analysis showed that the peripheral blood OPG value in the severe calcification group was significantly higher than mild calcification group (P=0.000) and non-calcification group (P=0.000), while the content of peripheral blood sRANKL in the severe calcification group was significantly lower than that in the non-calcification group (P=0.001). The valval OPG value in the mild calcification group was significantly higher than that of the non-calcification group (P=0.001) and the severe calcification group (P=0.040), and the valval sRANKL value of the severe calcification group was significantly lower than that of the non-calcification group (P=0.000).

Conclusion: The expression of OPG and RANKL can regulate the inflammatory reaction of valve and the balance between bone formation and resorption, thus affecting the progress of valve calcification.

Objective: We report the first documented case of concurrent ectopic complete hydatidiform mole (CHM) and high-grade serous carcinoma (HGSC) of the fallopian tube, associated with unique histologic features and mutations in the HGSC.

Case report: The patient presented with pelvic pain and vaginal bleeding. Laboratory examination revealed a positive urine pregnancy test and high serum beta-human chorionic gonadotropin (β-hCG). Transvaginal ultrasound demonstrated a left adnexal mass suspicious for ectopic pregnancy. Salpingectomy was performed, and the fallopian tube, noted to be ruptured with a visible ectopic pregnancy, demonstrated chorionic villi with diffuse hydropic enlargement and mild trophoblast hyperplasia. p57/Kip2 immunohistochemical staining (IHC) showed loss of expression in villous cytotrophoblasts and stromal cells, confirming CHM. An incidental 0.5 cm focus of HGSC was identified in the fallopian tube, associated with serous tubal intraepithelial carcinoma (STIC). The tumor exhibited solid, transitional cell carcinoma-like, and acinar patterns, with intraluminal mucin highlighted by Alcian blue and PAS-D stains. Patient underwent staging surgery which resulted in the finding of a 0.7 cm HGSC in the left ovary with morphology concordant to the tubal mass, except for a pseudo-endometrioid pattern in the ovary. Notably, the HGSC is positive (2+, 90%) for FOLR1 antigen and harbored a pathogenic mutation (p.R273H) in exon 8 of the TP53 gene.

Conclusion: This report emphasizes the crucial role of meticulous sampling and histopathologic examination of the fallopian tube, including the fimbriae, in all salpingectomy specimens. Furthermore, the case highlights the broad spectrum of morphologies encountered in HGSC, including mucinous differentiation. HGSC should be in the differential diagnosis when encountering mucin-producing high-grade carcinoma.

Objective: Morroniside (MOR) has been reported to ameliorate inflammation in cardiovascular and cerebrovascular disease; however, its impact and mechanism on rheumatoid arthritis (RA) remains unclear. This study aimed to investigate the beneficial role of morroniside in treating RA and explore the anti-invasive mechanism of morroniside on joint destruction.

Methods: In vitro (using primary rat articular fibroblast-like synoviocytes (FLSs)) a wound healing assay was used to detect the migration of primary rat FLSs. Quantitative RT-PCR was used to measure the transcription of matrix metalloproteinase (MMP) MMP2 and MMP9. Western blot was used to measure the expression of MMP2, MMP9, p65, phosphorylated-p65 (p-p65), inhibitor of nuclear factor (NF)-[Formula: see text]Bα (I[Formula: see text]Bα), I[Formula: see text]B kinase α/β (IKKα/β), and phosphorylated-IKKα/β. Immunofluorescence assay was used to measure the nuclear translocation of p65. In vivo (using rats with collagen-induced arthritis), the joint histopathological changes were detected by routine hematoxylin and eosin. Immunohistochemistry assay was used to measure the expression MMP2 and MMP9.

Results: Morroniside diminished tumor necrosis factor (TNF)α-stimulated migration of primary rat articular FLSs. Morroniside also attenuated RA-FLSs invasion into joint and joint destruction in rats with collagen-induced arthritis (CIA). Further analysis revealed that morroniside inhibited the overexpression of matrix metalloproteinase MMP2 and MMP9 in TNFα-stimulated primary rat articular FLSs and joints of CIA rats. Mechanistically, morroniside suppressed the activation of I[Formula: see text]B kinase α/β, which resulted in elevated levels of the inhibitor of nuclear factor (NF)-[Formula: see text]B.

Conclusion: The present study suggested that morroniside can prevent joint destruction by suppressing the activation of the NF-[Formula: see text]B/MMPs pathway, thereby preventing FLSs invasion.

Objective: To screen and identify potential biomarker for early rheumatoid arthritis (RA) by bioinformatic analysis and experimental investigation.

Methods: Transcriptome data of RA synovium was downloaded from GEO. Differentially expressed genes (DEGs), gene ontology (GO) functional annotation, and KEGG pathway were analyzed to inspect significative target genes. The protein-protein interaction was constructed using STRING database and Cytoscape to screen hub genes with least absolute shrinkage and selection operator (LASSO). The diagnostic effectivity of screened hub genes was analyzed with receiver operating characteristic (ROC). RA synovial fibroblast (SF) was treated with TNF-α (20ng/mL for 24h). RT-qPCR and Western blotting were used to measure mRNA and protein for screened hub gene.

Results: A total of 271 DEGs were found in GEO dataset. GO analysis indicated that DEGs mainly involved in phagocytosis, recognition and complement activation, etc. KEGG analysis suggested that DEGs were mostly enriched in the cytokine-cytokine receptor interaction, regulation of lipolysis in adipocytes, PPAR signaling pathway. LASSO regression and ROC curve indicated that ADIPOQ, CIDEA, FABP4, AQP7, LOC102723407, PLIN4, LIPE, CIDEC, PLIN1, and LEP had excellent diagnostic value. The area under ROC was 0.734. The level of ADIPOQ, LEP, LIPE, PLIN1, and PLIN4 were lower in RA group rather than that of control group (p<0.01). The higher expressions of CIDEC and FABP4 were found in RA group comparing to control group (p<0.001).

Conclusions: Identified hub genes might be valuable biomarkers for early RA diagnosis to promote precise and personal therapy.

Objective: Nrf2/ARE signaling pathway is reported to alleviate sleep anxiety in patients with sleep disorders. This study aimed at exploring the effect of escitalopram (ESC) on sleep anxiety in patients with cerebral small vessel disease (CSVD) and sleep disorder and its correlation with the Nrf2/ARE signaling pathway.

Methods: In the present study, we enrolled 80 CSVD patients (disease group) and 40 healthy patients (control group) in our hospital. The CSVD patients were classified into ESC treatment group and conventional treatment group and administered ESC and diazepam, respectively. After treatment, the patients' sleep quality was monitored within three months, and their symptoms were evaluated. Additionally, a mouse model of CSVD was set up, and the rats received intragastric administration of low, moderate, and high dosage of ESC or thymoquinone. The morphology of nerve cells and cognitive functions in the rat hippocampus were seen. TUNEL staining was conducted to detect nerve cell apoptosis and RT-qPCR, and Western blot analyses determined the expression levels of Nrf2 and ARE.

Results: Compared with conventional treatment group, the patients of ESC treatment group had shorter symptom improvement time and better sleep quality with a lower AHI and PSQI score. On the other hand, in the animal model, ESC treatment alleviated sleep disorders in CSVD rats, improved cognition and serum TNF-α levels, and protected nerve cells. Administration of ESC eliminated the expressions of Nrf2 and ARE and reduced nerve cell apoptosis dose-dependently. Moreover, Nrf2/ARE pathway activator (Danshensu) decreased rat sleep time and the level of serum TNF-α with more cell atrophy, while Nrf2/ARE pathway inhibitor (ML385) greatly improved sleep quality of CSVD rats.

Conclusion: ESC can effectively relieve sleep anxiety in patients with CSVD and sleep disorders through downregulating the Nrf2/ARE signaling pathway. ESC treatment increases patient's sleep time and serum TNF-α levels and attenuates nerve cell damage, further improving the patient's sleep quality.

Objective: We studied the role of LNCRNA 1197 in rheumatoid arthritis (RA) and its underlying mechanism.

Methods: We detected expression of LNCRNA 1197 in RA fibroblast-like synovial cells (RA-FLS) by RT-qPCR. Functional experiments were conducted to assess the impact of LNCRNA on cells as demonstrated by a Transwell assay. In addition, Western blot analysis was conducted to analyze the expression of proliferation-related proteins and flow cytometry and CCK-8 to analyze cell cycle and cell proliferation. We also identified the potential downstream target miRNA of LNCRNA 1197.

Results: LNCRNA 1197 was highly expressed in RA-FLS. When LNCRNA 1197 was knocked down, the cell proliferation, migration, and invasion of RA-FLS were alleviated, and pro-inflammatory cytokines were significantly downregulated. Interestingly, LNCRNA 1197 was indicated to target miR-206 and promoted progression of FLS by inhibiting miR-206.

Conclusion: Taken altogether, LNCRNA 1197 promotes the progression of RA-FLSs by miR-206 inhibition.