Mingli Gu

Systematic review and meta-analysis of flow cytometry in urinary tract infection screening.

BACKGROUND Automated urine sediment analysis of white blood cells (WBCs) and bacteria is a promising approach for urinary tract infections (UTIs) screening. However, available data on their screening efficacy is inconsistent. METHODS English articles from Pubmed, EMBASE, and Web of Science published before December 1, 2012 were analyzed. The Quality Assessment for Studies of Diagnostic Accuracy (QUADAS) tool was used to evaluate the quality of eligible studies. Performance characteristics of WBCs and bacteria (sensitivity, specificity, and other measures of accuracy) were pooled and examined by random-effects models. RESULTS Nineteen studies containing 22,305 samples were included. Pooled sensitivities were 0.87 (95% confidence interval [CI], 0.86-0.89) for WBCs and 0.92 (95% CI, 0.91-0.93) for bacteria. Corresponding pooled specificities were 0.67 (95% CI, 0.66-0.68) for WBCs and 0.60 (95% CI, 0.59-0.61) for bacteria. Areas under the summary receiver operating characteristics curves were 0.87 and 0.93 for WBCs and bacteria, respectively. The major limitation of eligible studies was that enrolled subjects were often not representative of clinical patient populations in which UTI would be suspected. CONCLUSIONS WBC and bacterial measurements by the UF-100 and UF-1000i are useful indicators in UTI screening; however, the performances of these systems should be rigorously evaluated by additional studies.

β-Arrestin 1 Modulates Functions of Autoimmune T Cells from Primary Biliary Cirrhosis Patients

ObjectivesPrimary biliary cirrhosis (PBC) is an autoimmune disease, characterized by antimitochondrial antibodies and autoreactive T cells causing destruction of the primary bile ducts. The molecular mechanisms regulating the autoreactive T cells remain elusive. β-Arrestins (βarr) are multifunctional signaling molecules that are crucial to T cell survival. We hypothesized that βarr plays a critical regulatory function in the autoreactive T cells of PBC patients.MethodsPatients with hepatic biliary cirrhosis (n = 60) were evaluated. Cytokine expression, T cell proliferation, and transcription factors were evaluated to assess regulatory functions in autoreactive T cells from the patient.ResultsOur studies showed that expression of βarr1 was elevated significantly in T lymphocytes from patients with PBC. Moreover, the level of βarr1 mRNA positively correlated with Mayo risk score in PBC patients. Based on modulation of βarr in autoreactive T cell lines, overexpression of βarr1 increased T cell proliferation, augmented interferon production, downregulated activities of nuclear factor κB and AP-1, promoted acetylation of histone H4 in the promoter regions of CD40L, LIGHT, IL-17 and interferon-γ, while downregulating acetylation of histone H4 in the promoter regions of TRAIL, Apo2, and HDAC7A, thereby regulating expression of these genes.ConclusionsOur findings suggest that βarr1 contributes to the pathogenesis of PBC, having significant implications for novel therapy strategy, further providing information for investigating the mechanisms of autoimmune disease.

The accuracy of glial fibrillary acidic protein in acute stroke differential diagnosis: A meta-analysis.

Abstract We performed a systematic review of English-language studies published during the past three decades to investigate the diagnostic performance of serum glial fibrillary acidic protein (GFAP) for the differential diagnosis of acute stroke, including intracerebral hemorrhage (ICH) and cerebral ischemia (CI). QUADAS tools were used to evaluate the quality of the study. Performance characteristics (diagnostic sensitivity, specificity, and other measures of accuracy) were pooled and examined using fixed-effects models. Four studies met the inclusion criteria, and included 109 patients with ICH and 381 patients with CI. The summary estimates for GFAP in the ICH diagnoses had a diagnostic sensitivity of 0.80 (95% confidence interval 0.71–0.88), a specificity of 0.97 (95% confidence interval, 0.94–0.98), and a diagnostic odds ratio (DOR) of 119.55 (95% confidence interval: 51.75–276.19). The area under curve (AUC) and Q value for the sROC curves were 0.97 and 0.92, respectively. Therefore, GFAP showed high diagnostic accuracy for acute stroke differential diagnosis.

Increased Siglec-1 Expression in Monocytes of Patients with Primary Biliary Cirrhosis

Objectives: To evaluate Siglec-1 protein (CD169) and mRNA levels in peripheral blood monocytes of patients with primary biliary cirrhosis (PBC) and investigate its role in PBC pathogenesis by looking for correlations between Siglec-1 expression and key PBC associated biochemical indices. Methods: FACS analysis was used to identify the percentage of peripheral blood monocytes positive for both CD14 and Siglec-1 in (a) 45 PBC patients, (b) 40 patients with liver cirrhosis after hepatitis B infection and (c) 36 healthy controls. Siglec-1 mRNA was measured by real-time RT-PCR and serum biomarkers by routine biochemistry. Results: The percentage of CD14-Siglec-1 double positive cells was significantly higher (p< 0.01) in PBC patients than in healthy controls or cirrhosis post-hepatitis patients (13.68 ± 2.44%, 1.0 ± 0.2 %, and 4.1 ± 0.5 %, respectively). Siglec-1 mRNA expression in the PBC group was 3.42 times higher than in healthy controls (p < 0.01). Conclusion: We investigated the role of Siglec-1 in PBC by assessing its expression in mononuclear cells of PBC patients and levels of secreted cytokines in cell supernatants after Siglec-1 RNA interference. It is possible that elevated Siglec-1 expression in peripheral blood monocytes of PBC patients is correlated with monocyte-mediated inflammatory responses during the development of PBC.

Resistance to activation-induced cell death and elevated FLIPL expression of CD4+ T cells in a polyI:C-induced primary biliary cirrhosis mouse model

Primary biliary cirrhosis (PBC) is a type of organ-specific autoimmune disease in which immune tolerance is impaired by an unknown mechanism. We established a PBC animal model by injecting C57BL/6 mice with polyI:C to study activation-induced cell death (AICD) in CD4+ T lymphocytes and changes of apoptosis-associated molecules as a first step to understand the immune tolerance of PBC mice. Obvious inflammatory cell infiltration was observed in the portal area of the liver tissues in model mice and antimitochondrial antibodies (AMA) positive rate was 80%. The AICD level in both splenic and hepatic CD4+ T cells in the model group were all lower than those in controls, and in the model group the level for hepatic CD4+ T cells were significantly lower than that for splenic CD4+ T cells. Quantitative PCR revealed that FasL mRNA and TRAIL expression in CD4+ T cells in the model group decreased significantly compared with that in the control group. Western blots revealed that the expression of the anti-apoptotic protein FLIPL in the model group increased significantly with the FLIPL expression in hepatic CD4+ T cells significantly higher than that in splenic CD4+ T cells. There was a positive linear correlation between the number of infiltrated portal areas and relative expression of FLIPL in splenic CD4+ T cells in model group. There were no obvious changes for caspase-8 in either group. These results show that the anti-apoptotic ability of CD4+ T lymphocytes play an important role in immune tolerance in the PBC mouse model, and elevated FLIPL expression may enhance this ability. The inhibition of FasL and TRAIL expression may also help enhance this anti-apoptotic ability in CD4+ T lymphocytes and contribute to the aggravation of portal area inflammation.

Increased Granulysin Expression in Peripheral Blood Cells of Patients with Primary Biliary Cirrhosis and Its Clinical Implications

IntroductionGranulysin is a cytotoxic molecule involved in cellular immune reactions.Materials and methodsThe levels of granulysin mRNA in the peripheral blood and serum granulysin of patients with primary biliary cirrhosis (PBC) were determined by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The expression of granulysin mRNA and serum protein in PBC was increased compared to the controls.ResultsThe expression of granulysin mRNA or serum protein showed close associations, respectively, with natural killer cell population in PBC patients. Serum granulysin was down-regulated by steroid and ursodeoxycholic therapy for PBC according to the improvement of severity of PBC. In addition, the expression of serum granulysin were related to serum total bilirubin and Mayo Clinic risk score. The serum granulysin reflected the cellular immune status of patients with PBC, and the expression correlated with the severity of PBC.ConclusionTherefore, there is a clinical benefit to monitoring granulysin as a biomarker of the prognosis of patients with PBC.

Decreased TLR2 signal expression in peripheral blood mononuclear cell from patients with cryptococcal meningitis

Toll‐like receptors are the most important pattern recognition receptors that can recognize conserved molecular structures shared by large groups of pathogens. Here, the aim was to determine the expression and role of TLR2 in peripheral blood mononuclear cells (PBMCs) from patients with cryptococcal meningitis and healthy controls. TLR2 expression was measured using RT‐PCR and western blotting. The role of TLR2 in cytokine production by PBMCs after Cryptococcus neoformans exposure was assessed in healthy controls prior to incubation with anti‐TLR2. TLR2 mRNA and protein expression were both weaker in patients with cryptococcal meningitis than in healthy controls. Furthermore, pre‐incubation of PBMCs from healthy donors with anti‐TLR2 led to reduced expression of IFN‐γ and IL‐12p70, but not of IL‐4 and IL‐10, following C. neoformans stimulation. Our results suggest that impaired expression of TLR2 may be involved in defective host defense to C. neoformans through an attenuated Th1 response.

BLyS and APRIL expression in peripheral blood mononuclear cells of cryptococcal meningitis patients and their clinical significance

OBJECTIVE To investigate the levels of APRIL, BlyS and receptors as TACI, BCMA and BAFF-R in peripheral blood mononuclear cells (PBMC) of cryptococcal meningitis (CM) patients and its clinical significance. METHODS PBMC from 30 CM patients and 32 healthy controls were isolated. The mRNA levels of APRIL, BLyS and BLyS receptors were detected by fluorescent quantitation PCR. The effect of PBMC from CM patients on in vitro growth of Cryptococcus neoformans was compared in presence and absence of BLyS. RESULTS PBMC of CM patients exhibited significantly lower BLyS, TACI and BCMA mRNA levels but significantly higher BAFF-R mRNA levels than controls. Growth of C. neoformans was significantly slower in presence of BLyS than its absence. CONCLUSION Levels of BlyS and its receptors correlated with cryptococcal meningitis progression, and provide new clues for monitoring CM conditions and its effective therapy.

Lower Plasma Levels of IL-35 in Patients with Primary Biliary Cirrhosis.

Primary biliary cirrhosis (PBC) is an autoimmune liver disease. Its histological characteristics, such as progressive intrahepatic bile duct destruction, cholestasis, and liver cirrhosis, are caused by the bodys autoimmune disorders. Interleukin (IL)-35 has two subunits (p35 and Ebi3) and is a member of the IL-12 family of heterodimeric cytokines. IL-35 has immunosuppressive functions and plays an important role in many autoimmune diseases. In this study, we compared plasma levels of IL-35 and relative mRNA expression levels of p35 and Ebi3 in peripheral blood mononuclear cells (PBMCs) from 70 PBC patients and 70 healthy individuals. The results showed that the relative expression levels of Ebi3 mRNA were lower in PBMCs from PBC patients than in PBMCs from healthy individuals, whereas the levels of p35 mRNA were similar in both groups. Plasma IL-35 concentrations were lower in patients with PBC than in healthy individuals. Plasma levels were higher in PBC patients at an advanced stage compared to patients at an early stage. Variable plasma levels with different stages were also found in transforming growth factor beta (TGF-β), which is mainly produced by regulatory T cells (Tregs). IL-35 and TGF-β levels were positively correlated with each other, and IL-35 was capable of promoting the inhibitory functions of Tregs in PBC patients at both the early and late stages of disease. Lower plasma IL-35 levels were accompanied by higher levels of typical clinical parameters, such as alkaline phosphatase, or of proinflammatory cytokines, such as interferon-gamma (IFN-γ), in PBC patients (P < 0.05 for each). We propose that IL-35 may be involved in the pathogenesis of PBC and could be a potential biomarker for diagnosing this disease.

Aberrant IL-35 levels in patients with primary Sjogren's syndrome

IL‐35 is a newly discovered immunoregulatory cytokine that possesses the ability to inhibit CD4 + effector T cells and alleviate autoimmune diseases. The objective of this study was to investigate IL‐35 levels in patients with primary Sjogrens syndrome (pSS) and explore the roles of IL‐35 in the pathogenesis of pSS.