Aberrant alteration of peripheral B lymphocyte subsets in hepatocellular carcinoma patients.

Although B lymphocytes are widely known to participate in the immune response, the conclusive roles of B lymphocyte subsets in the antitumor immune response have not yet been determined. Single-cell data from GEO datasets were first analyzed, and then a B cell flow cytometry panel was used to analyze the peripheral blood of 89 HCC patients and 33 healthy controls recruited to participate in our research. Patients with HCC had a higher frequency of B10 cells and a lower percentage of MZB cells than healthy controls. And the changes in B cell subsets might occur at an early stage. Moreover, the frequency of B10 cells decreased after surgery. Positively correlated with B10 cells, the elevated IL-10 level in HCC serum may be a new biomarker in HCC identification. For the first time, our results suggest that altered B cell subsets are associated with the development and prognosis of HCC. Increased B10 cell percentage and IL-10 in HCC patients suggest they might augment the development of liver tumors. Hence, B cell subsets and related cytokines may have predictive value in HCC patients and could be potential targets for immunotherapy in HCC.

[Carrier screening for spinal muscular atrophy in 4719 pregnant women in Shanghai region].

OBJECTIVE: Spinal muscular atrophy (SMA) is a common and fatal autosomal recessive disorder. Approximately 94% of SMA patients are caused by homozygous deletion of SMN1 gene. SMA carrier screening is recommended considering the high carrier frequency (1 in 35-50) as well as severity of the disease. METHODS: A prospective population-based cohort study was carried out on 4719 pregnant women from Shanghai region. Copy numbers of SMN1 and SMN2 genes were effectively determined with denaturing high performance liquid chromatography (DHPLC) technique. The method has detected 94% of SMA cases with deletion or conversion of the SMN1 genes. RESULTS: Ninety SMA carriers with only one copy of the SMN1 gene were identified among the 4719 pregnant woman. The carrier rate was 1.9%. Respectively, 1.2% and 0.6% of the carriers were caused by SMN1 gene deletion and SMN1 gene conversion. CONCLUSION: Through this study, we have determined the frequency of SMA mutation carriers in a population of pregnant women. The result may provide a basis for genetic counseling in order to reduce the rate of SMA affected births.

GeneXpert MTB/RIF combined with conventional methods for tuberculosis in Shanghai Regional Medical Center: a retrospective diagnostic study.

Background: At present, the diagnosis of tuberculosis (TB) is still challenging, and improving the efficiency of diagnosis can help prevent and control TB. This retrospective clinical study aimed to assess the diagnostic efficiency of GeneXpert MTB/RIF for pulmonary TB. Methods: A total of 620 newly-diagnosed patients who visited the pulmonary clinic of Shanghai Tongren Hospital between 2018 and 2021 were enrolled in the study. All 620 patients had acid-fast Bacilli (AFB) identified by Ziehl Neelsen staining (ZNS) test, BECTEC MGIT 960 liquid culture (LC), and GeneXpert MTB/RIF assay (GX). A total of 53 patients also underwent interferon-γ release assay (IGRA). The diagnostic efficacy of ZNS, LC, GX alone or in combination in pulmonary TB was evaluated, with clinical diagnosis as the gold standard. Moreover, the IGRA for pulmonary TB diagnosis was preliminarily assessed. Results: Eventually, 185 cases were clinically confirmed (which included 36 etiologically negative cases) in the total enrolled 620 first-diagnosed patients. Overall, the 3 methods ZNS, LC, and GX showed sensitivities of 55.68%, 64.32%, and 68.64%, specificities of 98.39%, 95.40%, and 99.08%, positive predictive values (PPV) of 93.64%, 85.61%, and 96.95%, and negative predictive values (NPV) of 83.92%, 86.28%, and 88.14%, respectively. The GX method showed the highest specificity and PPV for a solitary single method, with 99.08% and 96.95%, respectively. Regarding pairwise combination methods, all showed superior sensitivity to a single test, reaching a maximum of 80.00%. Among them, the LC + GX combination showed both the highest sensitivity (80.00%) and NPV (91.78%), and the corresponding area under the receiver operating characteristic curve (0.875) was the largest. Among the 53 patients who underwent IGRA testing, 42 were positive (including 4 etiologically negative cases), and 11 were negative. The overall sensitivity of IGRA for diagnosing pulmonary TB was 90.00%, specificity was 27.27%, PPV was 42.86%, and NPV was 81.82%. Conclusions: The GX method shows promise as a first-line diagnostic method for pulmonary TB. Furthermore, the sensitivity was significantly improved when combined with LC. This combination will screen out some etiologically negative patients plus IGRA, so their combination is recommended for practice optimization.

m6A RNA methylation-mediated NDUFA4 promotes cell proliferation and metabolism in gastric cancer.

Gastric cancer (GC) is a malignancy with poor prognosis. NDUFA4 is reported to correlate with the progression of GC. However, its underlying mechanism in GC is unknown. Our study was to reveal the pathogenic mechanism of NDUFA4 in GC. NDUFA4 expression was explored in single-cell and bulk RNA-seq data as well as GC tissue microarray. Mitochondrial respiration and glycolysis were estimated by oxygen consumption rate and extracellular acidification rate, respectively. The interaction between NDUFA4 and METTL3 was validated by RNA immunoprecipitation. Flow cytometry was used to estimate cell cycle, apoptosis and mitochondrial activities. NDUFA4 was highly expressed in GC and its high expression indicated a poor prognosis. The knockdown of NDUFA4 could reduce cell proliferation and inhibit tumor growth. Meanwhile, NDUFA4 could promote glycolytic and oxidative metabolism in GC cells, whereas the inhibition of glycolysis suppressed the proliferation and tumor growth of GC. Besides, NDUFA4 inhibited ROS level and promoted MMP level in GC cells, whereas the inhibition of mitochondrial fission could reverse NDUFA4-induced glycolytic and oxidative metabolism and tumor growth of GC. Additionally, METTL3 could increase the m6A level of NDUFA4 mRNA via the m6A reader IGF2BP1 to promote NDUFA4 expression in GC cells. Our study revealed that NDUFA4 was increased by m6A methylation and could promote GC development via enhancing cell glycolysis and mitochondrial fission. NDUFA4 was a potential target for GC treatment.

Interaction between early-life pet exposure and methylation pattern of ADAM33 on allergic rhinitis among children aged 3-6 years in China.

BACKGROUND: Recent research has pointed out the important roles of epigenetic modifications in the development and persistence of allergic rhinitis (AR), especially in relation to DNA methylation of disease-associated genes. We investigated whether AR susceptibility genes were epigenetically regulated, and whether methylation modulation of these genes in response to early-life environment could be a molecular mechanism underlying the risk for AR onset in a cohort of children aged 3-6 years in China. METHODS: Peripheral blood mononuclear cell (PBMC) samples were collected from 130 children patients, aged 3-6 years and diagnosed with AR; and 154 matched controls to detect promoter methylation in 25 AR susceptibility genes with the MethylTarget approach. Methylation levels were compared for each CpG site, each amplified region, and each gene. In addition, the relationship among DNA methylation, early-life environmental risk factors and AR onset were assessed. RESULTS: Maternal allergic history (P = 0.0390) and pet exposure (P = 0.0339) were significantly associated with increased AR risk. Differential methylation analyses were successfully performed for 507 CpG sites, 34 amplified regions and 17 genes and significant hypomethylation was observed in the promoter region of ADAM33 in AR patients [multiple test-corrected (FDR) P-value < 0.05]. Spearman correlation analysis revealed that the hypomethylation of ADAM33 was significantly associated with higher eosinophil counts (Spearman's ρ: - 0.187, P-value = 0.037). According to the results of the multiple regression analysis, after adjusting for cofounders, the interaction of early-life pet exposure with methylation level of ADAM33 increased the risk for AR onset 1.423 times more in children (95% CI = 0.0290-4.109, P-value = 0.005). CONCLUSION: This study provides evidence that early-life pet exposure and low methylation level of ADAM33 increase AR risk in children, and the interaction between pet exposure and methylation level of ADAM33 may play an important role in the development of AR.

Leptin receptor mediates the proliferation and glucose metabolism of pancreatic cancer cells via AKT pathway activation.

Pancreatic cancer (PC) is the fourth leading cause of cancer­related mortality worldwide. Leptin is an adipokine that is significantly increased in obese patients and that functions in various biological processes of cancer, such as tumor growth and metastasis. However, its role in PC cell proliferation and glucose metabolism and the underlying mechanisms remain unclear. In the present study, in vitro leptin treatment significantly promoted cell proliferation and increased glucose uptake and lactate production of human PC and healthy pancreas cells in a dose­dependent manner, accompanied by increased expression of the glycolytic enzymes hexokinase II and glucose transporter 1. Furthermore, leptin receptor­specific short hairpin RNAs were used to silence leptin receptor expression in PC cells, which had the opposite effect to leptin stimulation and decreased AKT phosphorylation. In addition, the effects of leptin stimulation were significantly counteracted by the AKT inhibitor LY294002, whereas the effects of leptin silencing were counteracted by AKT activator insulin­like growth factor 1. The results of the present study suggested that leptin may contribute to cell proliferation and glucose metabolism of human PC cells, which may be through activation of the AKT pathway.

Validation of the BD FACSPresto system for the measurement of CD4 T-lymphocytes and hemoglobin concentration in HIV-negative and HIV-positive subjects.

This study aimed to compare the performance of the BD FACSPresto system with the conventional standard-of-care technologies for the measurement of absolute CD4 count (AbsCD4), CD4 percentage (CD4%) and total hemoglobin concentration (Hb) in capillary and venous blood samples of HIV-negative and HIV-positive subjects. A total of 1304 participants were included in this prospective cohort study. Both venous and capillary blood samples were analyzed using the BD FACSPresto system and the results were compared against the BD FACSCalibur for enumerating AbsCD4 and CD4% and Sysmex XT-4000i hematology analyzer for determining Hb levels. Method comparison studies were performed using Deming regression and Bland-Altman plots. The Deming regression analyses comparing the accuracy of the BD FACSPresto system with the reference standard technologies demonstrated a significant linear correlation between the AbsCD4, CD4%, and Hb values generated by the two platforms. The 95% CI of the slopes for AbsCD4, CD4%, and Hb levels were 0.94-0.99, 0.99-1.01 and 0.86-0.93, respectively (P < 0.001). Bland-Altman plots for AbsCD4, CD4%, and Hb levels demonstrated close agreement between the BD FACSPresto system and the reference standards for all study participants. The performance and accuracy of BD FACSPresto system was comparable to the reference standard technologies. The BD FACSPresto system can be used interchangeably with BD FACSCalibur platform for CD4 and Sysmex XT-4000i hematology analyzer for Hb concentrations in resource-limited settings thus, improving accessibility to point-of-care testing services.