Diagnostic Utility of CD64 and CD38 Biomarkers for the Differential Diagnosis of Infections.

Purpose: Identification of infection type in patients with fever is particularly important in the emergency departments (EDs) of hospitals. This study was designed to evaluate the performance of two biomarkers, the modified neutrophil CD64 (nCD64) index and CD38 presence on T cells, using flow cytometry. Methods: A total of 305 potentially infected patients with fever were admitted to the ED of Zhongda Hospital (Nanjing, China) between March 2021 and August 2021. This study included three groups of patients: bacterial (N = 180), viral (N = 30) , and uninfected (N = 65) based on their final diagnostic outcomes and clinical records. Results: The expression level of traditional/modified nCD64 was significantly increased in the bacterial infection group, especially in case of patients infected with Gram-negative bacteria. The most prevalent species were Staphylococcus spp. and Escherichia coli. In contrast, CD3+CD38+ cell percentages were elevated in patients with viral infections, which were mostly caused by Epstein-Barr virus and cytomegalovirus. CD38 expression is age dependent, and higher percentages of CD3+CD38+ cells were observed in children with viral infections. For the prediction of bacterial infections, the area under the curve (AUC) of modified nCD64 (AUC: 0.800) was significantly higher than that of C-reactive protein and heparin-binding protein but slightly lower than that of traditional nCD64 (AUC: 0.831). The AUC, specificity, and sensitivity values for the prediction of viral infections using CD3+CD38+ cells percentages in children were 0.899 (0.785-1.000), 96.2%, and 85.9%, respectively. Conclusion: nCD64 levels and CD3+CD38+ cell percentage are potential biomarkers that facilitate identification of patients with bacterial and viral infections.

Activity of the antimicrobial peptide and thanatin analog S-thanatin on clinical isolates of Klebsiella pneumoniae resistant to conventional antibiotics with different structures.

The treatment of infections caused by bacteria resistant to the vast majority of antibiotics is a challenge worldwide. To evaluate the effect of S-thanatin (an analog of thanatin, a cationic antimicrobial peptide isolated from the hemipteran insect Podisus maculiventris) against microbial resistant to antibiotics, we studied its bactericidal kinetics, synergistic effect, resistance, and activity on clinical isolates of Klebsiella pneumoniae resistant to conventional antibiotics with different structures. The bactericidal rate of S-thanatin was more than 99% against K. pneumoniae ATCC 700603 when bacterial cultures were monitored for 60 min. The peptide was synergistic with beta-lactam cefepime in most of the clinical MDR isolates tested (7/8). An average value of FIC was 0.3708. No synergy was found between the peptide and amoxicillin, gentamycin, tetracycline, or ciprofloxacin in all bacteria tested. A total of 48 isolates of K. pneumoniae with different resistance spectrum tested was susceptible to S-thanatin. The MICs were 6.25-25 mug/ml. No significant difference in the MICs of S-thanatin between the sensitive isolates and the resistant isolates to single antibiotic was observed (P > 0.05). The resistance of K. pneumoniae ATCC 700603 to S-thanatin was slightly higher, when cultured at sub-inhibitory concentration for 5 days. S-thanatin may be an attractive candidate for developing into an antimicrobial agent.

Role of connective tissue growth factor (CTGF) module 4 in regulating epithelial mesenchymal transition (EMT) in HK-2 cells.

BACKGROUND: Recent studies have suggested that connective tissue growth factor (CTGF) plays a key role in tissue fibrosis including renal scarring. While studies showed several forms of CTGF with 10-38 kDa in the body fluids, little is known about these small molecule species. We investigated the effect of a 10 kDa CTGF molecule consisting of module 4, on the epithelial mesenchymal transition (EMT) in human proximal tubular cell line (HK-2). METHODS: HK2 cells were cultured in DMEM medium. The response of cytokeratin (CK) and vimentin (VIM) mRNA and protein expression to the stimulation of rhCTGF(C) were observed by real-time PCR and immunocytochemistry. At the same time, the morphologic changes were observed by microscopy, and expression of alpha-smooth muscle actin (alpha-SMA) and fibronectin (FN) was detected by laser confocal microscope. These effects were compared with CTGF N-terminal [rhCTGF(N)], consisting of module 1-3, and observed in a condition with the addition of anti-CTGF antibody. RESULTS: RhCTGF(C) induced striking changes in epithelial cells, including changes in cellular morphology, loss of CK, gain VIM and alpha-SMA, and increased levels of fibronectin. Cocultured with anti-CTGF antibody could abrogate most of these effects, while cells treated with rhCTGF(N) showed no significant phenotypic changes comparing to control group. CONCLUSIONS: Our results suggest that module 4 could induce HK-2 cells EMT, whereas the residual fragment has no similar effect in spite of consisting of 3 modules of CTGF molecule.

Multiplex real-time PCR for RRM1, XRCC1, TUBB3 and TS mRNA for prediction of response of non-small cell lung cancer to chemoradiotherapy.

BACKGROUND: This study was aimed to establish a novel method to simultaneously detect expression of four genes, ribonucleotide reductase subunit M1(RRM1), X-ray repair cross-complementing gene 1 (XRCC1), thymidylate synthase (TS) and class III ß-tubulin (TUBB3), and to assess their application in the clinic for prediction of response of non-small cell lung cancer (NSCLC) to chemoradiotherapy. MATERIALS AND METHODS: We have designed four gene molecular beacon (MB) probes for multiplex quantitative real-time polymerase chain reactions to examine RRM1, XRCC1, TUBB3 and TS mRNA expression in paraffin-embedded specimens from 50 patients with advanced or metastatic carcinomas. Twenty one NSCLC patients receiving cisplatin- based first-line treatment were analyzed. RESULTS: These molecular beacon probes could specially bind to their target genes in homogeneous solutions. Patients with low RRM1 and XRCC1 mRNA levels were found to have apparently higher response rates to chemoradiotherapy compared with those with high levels of RRM1 and XRCC1 expression (p<0.05). The TS gene expression level was not significantly associated with chemotherapy response (p>0.05). CONCLUSIONS: A method of simultaneously detecting four molecular markers was successfully established and applied for evaluation of chemoradiotherapy response. It may be a useful tool in personalized cancer therapy.

Tissue kallikrein is related to the severity of coronary artery disease.

BACKGROUND: The impairment of the tissue kallikrein (KLK1)-kinin system (KKS) may result in atheroma development. However, it remains unclear if the KKS correlates with coronary artery disease (CAD). METHODS: KLK1, VEGF and hs-CRP plasma levels were measured in 100 patients newly diagnosed with CAD and 33 CAD-free controls. Patients were followed-up for the incidence of major adverse cardiovascular events (MACE) for 8months to 2y. Gene expression of KLK1, CD105 and CD68 was assessed in human coronary endarterectomy specimens. RESULTS: Patients with CAD and acute coronary syndrome (ACS) had significantly elevated KLK1 levels. In addition, the concentration of hs-CRP was increased in ACS patients. A strong positive correlation between plasma KLK1 and the severity of CAD was also demonstrated, suggesting that high KLK1 levels are an independent predictor for CAD. MACE during follow-up significantly correlated with KLK1 levels in the ACS group. Unstable coronary plaques demonstrated markedly increased KLK1 levels, macrophage infiltration and high microvessel density. Additionally, KLK1 staining primarily colocalized with macrophages. CONCLUSIONS: In the present study, plasma KLK1 levels were a useful predictor for the presence and extent of CAD. More extensive studies are, however, necessary in order to validate these findings.