Diagnostic performance and prognostic value of circulating tumor DNA methylation marker in extranodal natural killer/T cell lymphoma.

Circulating tumor DNA (ctDNA) carries tumor-specific genetic and epigenetic variations. To identify extranodal natural killer/T cell lymphoma (ENKTL)-specific methylation markers and establish a diagnostic and prognosis prediction model for ENKTL, we describe the ENKTL-specific ctDNA methylation patterns by analyzing the methylation profiles of ENKTL plasma samples. We construct a diagnostic prediction model based on ctDNA methylation markers with both high specificity and sensitivity and close relevance to tumor staging and therapeutic response. Subsequently, we built a prognostic prediction model showing excellent performance, and its predictive accuracy is significantly better than the Ann Arbor staging and prognostic index of natural killer lymphoma (PINK) risk system. Notably, we further establish a PINK-C risk grading system to select individualized treatment for patients with different prognostic risks. In conclusion, these results suggest that ctDNA methylation markers are of great value in diagnosis, monitoring, and prognosis, which might have implications for clinical decision-making of patients with ENKTL.

Preparation of artificial antigen and egg yolk-derived immunoglobulin (IgY) of citrinin for enzyme-linked immunosorbent assay.

OBJECTIVE: To prepare artificial antigens and anti-citrinin egg yolk-derived immunoglobulin (IgY) to build an enzyme-linked immunosorbent assay (ELISA) for citrinin (CTN). METHODS: CTN was conjugated with bovine serum albumin (BSA), ovalbumin (OVA) with formaldehyde condensation method to prepare artificial antigens and identified by ultraviolet (UV) spectrometry and Infrared (IR) spectrometry. Artificial antigens for CTN and anti-CTN IgY were purified with polyethylene glycol two-step precipitation method and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). ELISA with IgY was established. Cross-reactivity of IgY with various structural similarities to CTN and possible co-occurrence with CTN in agricultural commodities were studied. RESULTS: UV and IR absorption spectra suggested that CTN was correlated with the carrier protein of BSA or OVA. SDS-PAGE patterns showed that the anti-CTN IgY was almost pure with a molecular weight of approximate 100 KD. The indirect competitive ELISA showed that the detection limit of CTN was 10 ng x mL(-1), with a good linearity ranging 20-640 ng x mL(-1). CONCLUSION: Artificial antigens of CTN can be successfully synthesized. The established ELISA can be used to determine CTN- contaminated samples.

Serum neopterin for early assessment of severity of severe acute respiratory syndrome.

Neopterin and C-reactive protein (CRP) concentrations were determined in serum samples from 129 severe acute respiratory syndrome (SARS) patients and 156 healthy blood donors. In the patients with confirmed SARS, an early neopterin elevation was detected already at the day of onset of symptoms and rose to a maximum level of 45.0 nmol/L 3 days after the onset. All SARS patients had elevated neopterin concentrations (>10 nmol/L) within 9 days after the onset. The mean neopterin concentrations were 34.2 nmol/L in acute sera of SARS patients, 5.1 nmol/L in convalescent sera, and 6.7 nmol/L in healthy controls. In contrast, the mean CRP concentrations in both acute and convalescent sera of SARS patients were in the normal range (<10 mg/L). Serum neopterin level in SARS patients was associated with fever period and thus the clinical progression of the disease, while there was no significant correlation between the CRP level and the fever period. Serum neopterin may allow early assessment of the severity of SARS. The decrease of neopterin level was found after steroid treatment, which indicates that blood samples should be collected before steroid treatment for the neopterin measurement.

Th2 predominance and CD8+ memory T cell depletion in patients with severe acute respiratory syndrome.

UNLABELLED: The immune spectrum of severe acute respiratory syndrome (SARS) is poorly understood. To define the dynamics of the immune spectrum in SARS, serum levels of cytokines, chemokines, immunoglobulins, complement and specific antibodies against SARS-associated coronavirus (SARS-CoV) were assayed by enzyme-linked immunosorbent assay (ELISA), and phenotypes of peripheral lymphocytes were analyzed by flow cytometry in 95 SARS-infected patients. Results showed that interleukin (IL)-10 and transforming growth factor beta (TGF-beta) were continuously up-regulated during the entirety of SARS. Regulated on activation normally T cell-expressed and secreted (RANTES) levels were decreased, while monocyte chemoattractant protein-1 (MCP-1) was elevated in acute patients. Immunoglobulins and complement were elevated during the first month of SARS. Both serum-positive rates and titers of specific IgM and IgG antibodies responding to SARS-CoV peaked at days 41-60 from the onset of SARS. CD4+ and CD8+ T lymphocytes decreased significantly in acute-phase. CD3+CD8+CD45RO+ T lymphocytes were decreased by 36.78% in the convalescent patients. CONCLUSION: SARS-CoV seemed to elicit effective humoral immunity but inhibited cellular immunity, especially CD8+ memory T lymphocytes over time. Prolonged overproduction of IL-10 and TGF-beta may play an important role in the disease.

[The value of Roferon-A combined with hepatic artery chemoembolization and portal vein chemotherapy after radical resection in hepatocellular carcinoma for preventing recurrence].

OBJECTIVE: To explore the change of T cell subsets in patients suffered from hepatocellular carcinoma (HCC) before and after hepatectomy, and study the value of Roferon-A (interferon alpha-2a) combined with hepatic artery chemoembolization (HACE) and portal vein chemotherapy (PVC) after radical resection of HCC for preventing recurrence. METHODS: On 75 HCC patients, PVC and HACE were respectively given at 2 weeks and 4 weeks after radical tumor resection. In 2nd week after surgery, 33 cases of them accepted Roferon-A treatment for 1 week. Seventy-two patients were followed up over 3 years. Effect of Roferon-A combined with HACE and PVC on postoperative recurrence rate was compared with that of HACE and PVC. Changes of T cell subsets in peripheral blood were examined with labeled monoclonal antibodies before and after hepatectomy or using interferon. Forty cholecystolithiasis patients received cholecystectomy were used as the controls. RESULTS: CD(3)(+) and CD(4)(+) cells in peripheral blood were reduced in patients with HCC. After hepatectomy, they declined further with decrease in CD(4)(+)/CD(8)(+) ratio. The results returned to pre-operative level at the end of 4th week after surgery. The CD(3)(+), CD(4)(+) cells and the CD(4)(+)/CD(8)(+) ratio increased remarkably following the use of Roferon-A. The 1-, 2- and 3-year recurrence rates of patients treated with HACE, PVC and Roferon-A in combination were 0%, 6.2% and 15.6%, respectively, while those treated with HACE and PVC were 5.0%, 12.5% and 27.5%, respectively. CONCLUSION: Patients with HCC suffer from marked immuno-suppression which became ever more severe after hepatectomy, combined use of HACE, PVC and Roferon-A is superior to only HACE and PVC by decreasing the recurrence rate.