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Hand, foot and mouth disease (HFMD) was one of the most common infectious disease in the past few decades. After 2013, coxsackievirus A6 (CVA6) has replaced enterovirus 71 (EV-A71) and coxsackievirus A16 (CVA16), becoming the predominant pathogen responsible for HFMD in many areas in China. The objective of this study is to investigate the genetic characteristics and molecular epidemiology of CVA6 in Linyi from 2022 to 2023. A total of 965 HFMD cases were enrolled in this study and analyses based on VP1 nucleotide sequences were performed to determine the evolutionary trajectory of CVA6. In 2022, 281 (281/386, 72.8%) were positive for enterovirus (EVs), and 217 (217/281, 77.2%) were CVA6 positive. In 2023, 398 (398/579, 68.7%) samples were positive for EVs, and 243 (243/398, 61.1%) were CVA6 positive. Six sequences were selected from each year for the homology analysis. The results showed that 12 strains isolated in Linyi were far from the prototype strain (AY421764) and the first CVA6 strain reported in China (JQ364886). Phylogenetic analysis showed that the CVA6 strains isolated in Linyi all belonged to D3 subgenotype. CVA6 is emerging as a common pathogen causing HFMD in Linyi, and continuous surveillance of HFMD etiological agents is necessary.

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Serum magnesium levels exceeding 0.9 mmol/L are associated with increased survival rates in patients with CKD. This retrospective study aimed to identify risk factors for cardio-cerebrovascular events among patients receiving continuous ambulatory peritoneal dialysis (CAPD) and to examine their correlations with serum magnesium levels. Sociodemographic data, clinical physiological and biochemical indexes, and cardio-cerebrovascular event data were collected from 189 patients undergoing CAPD. Risk factors associated with cardio-cerebrovascular events were identified by univariate binary logistic regression analysis. Correlations between the risk factors and serum magnesium levels were determined by correlation analysis. Univariate regression analysis identified age, C-reactive protein (CRP), red cell volume distribution width standard deviation, red cell volume distribution width corpuscular volume, serum albumin, serum potassium, serum sodium, serum chlorine, serum magnesium, and serum uric acid as risk factors for cardio-cerebrovascular events. Among them, serum magnesium ≤0.8 mmol/L had the highest odds ratio (3.996). Multivariate regression analysis revealed that serum magnesium was an independent risk factor, while serum UA (<440 µmol/L) was an independent protective factor for cardio-cerebrovascular events. The incidence of cardio-cerebrovascular events differed significantly among patients with different grades of serum magnesium (χ2 = 12.023, p = 0.002), with the highest incidence observed in patients with a serum magnesium concentration <0.8 mmol/L. High serum magnesium levels were correlated with high levels of serum albumin (r = 0.399, p < 0.001), serum potassium (r = 0.423, p < 0.001), and serum uric acid (r = 0.411, p < 0.001), and low levels of CRP (r = -0.279, p < 0.001). In conclusion, low serum magnesium may predict cardio-cerebrovascular events in patients receiving CAPD.

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A high-precision binocular camera calibration method is proposed to address the issues of poor calibration accuracy and large calibration errors in current practical applications. This method uses a triangular stereo sphere as the calibration object and employs steps, such as ellipse fitting, Cholesky decomposition, homography matrix solution, and nonlinear optimization, to compute the intrinsic and extrinsic parameters, distortion parameters, and relative pose of the binocular camera. Moreover, this method simplifies the correspondences between primitives, enabling simultaneous calibration of multiple viewpoint cameras. This method is also suitable for both binocular cameras consisting of two different structured monocular cameras and those composed of two image sensors within the same structure. Experimental results showed that this method outperforms traditional algorithms in terms of binocular camera calibration accuracy, calibration errors between left and right cameras, and robustness, resulting in a significant improvement in overall algorithm performance.

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Membranous nephropathy (MN) is an autoimmune disease characterized by the deposition of immunoglobulin G (IgG) and complementary components in the epithelium of the glomerular capillary wall. Macrophage migration inhibitory factor (MIF) is an inflammatory mediator released by macrophages. MIF plays a key regulatory function in the pathogenesis of immune-mediated glomerulonephritis. This study aimed to investigate whether MIF level could be associated with the activity of MN. Plasma and urine samples from 57 MN patients and 20 healthy controls were collected. The MIF levels in plasma and urine were determined by an enzyme-linked immunosorbent assay (ELISA) kit. The expression of MIF in the renal specimens from 5 MN patients was detected by immunohistochemistry (IHC). The associations of the plasma and urinary levels of MIF and glomerular MIF expression with clinical and pathological characteristics were analyzed. It was revealed that with the increase of MIF levels in plasma and urine, the severity of renal pathological injury in MN patients gradually increased. Correlation analysis showed that the MIF levels in plasma were positively correlated with the platelet (PLT) count (r = 0.302, P = 0.022), and inversely correlated with the prothrombin time (PT) (r = - 0.292, P = 0.028) in MN patients. The MIF levels in plasma were positively correlated with the C-reactive protein (CRP) level and erythrocyte sedimentation rate (ESR) (r = 0.651, P < 0.0001; r = 0.669, P < 0.0001) in MN patients. The urinary levels of MIF were positively correlated with ESR (r = 0.562, P < 0.0001). IHC suggested that MIF was expressed in glomerular basement membrane and tubulointerstitial areas. MIF levels in plasma and urine could reflect the severity of MN, and MIF levels in plasma and urine could be associated with venous thrombosis and infectious complications in MN patients. The glomerular MIF expression could be used to indicate the activity of MN.

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BACKGROUND: C5a is important for antineutrophil cytoplasmic antibody (ANCA)-mediated activation of neutrophils. The present study aimed to assess the role of macrophage migration inhibitory factor (MIF) in ANCA-mediated activation of C5a-primed neutrophils. The effects of MIF on ANCA-mediated neutrophil respiratory burst and degranulation were determined. In addition, the effect of a MIF antagonist on the activation of C5a-primed neutrophils was assessed. RESULTS: MIF treatment resulted in increased membrane proteinase-3 (mPR3) expression on neutrophils and enhanced myeloperoxidase (MPO) amounts in neutrophil culture supernatants. The concentration of MIF was significantly higher in the neutrophils supernatant primed with C5a (negative control: 14.2 ± 1.16 ng/ml; C5a: 45.8 ± 2.8 ng/ml, P < 0.001 vs. negative control; C5a + IgG: 44.8 ± 1.93 ng/ml, P < 0.001 vs. negative control; C5a + MPO-ANCA: 73.0 ± 5.5 ng/ml, P < 0.001 vs. C5a; and C5a + PR3-ANCA: 69.4 ± 5.35 ng/ml, P < 0.001 vs. C5a). MIF primed neutrophils to undergo respiratory burst and degranulation in response to ANCA. Indeed, mean fluorescence intensity (a measure of respiratory burst) was significantly higher in MIF-primed neutrophils activated with MPO-ANCA-positive IgG or PR3-ANCA-positive IgG compared with non-primed neutrophils. Meanwhile, a MIF antagonist reduced oxygen radical production in C5a-primed neutrophils treated with patient-derived ANCA-positive IgG. CONCLUSIONS: MIF can prime neutrophils to undergo ANCA-mediated respiratory burst and degranulation. Blocking MIF resulted in reduced ANCA-mediated activation of C5a-primed neutrophils. These findings indicated that the interaction between MIF and C5a may contribute to ANCA-mediated neutrophil activation.

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Atherosclerotic diseases are systemic and patient outcomes depend on comprehensive imaging evaluation. Computed tomography angiography (CTA) is a powerful tool used to assess atherosclerosis. However, the scanning protocol is designed for cardiovascular and cerebrovascular imaging, which require considerations into the radiation dose, contrast agent and image quality. The purpose of the present study was to evaluate ultra-low dose one-step CTA for coronary, carotid and cerebral arteries with a low concentration contrast agent. A total of 78 patients were enrolled and randomly divided into two groups: Group A (n=38) and B (n=40). High-pitch CTA for coronary, carotid and cerebral arteries with a tube voltage of 70 or 80 kVp and 40 ml contrast agent (270 mgI/ml) was performed by a 128-slice dual-source CT scanner for group A. Standard high-pitch CTA with a tube voltage of 100 kVp and 60 ml contrast agent (370 mgI/ml) was conducted for group B. The image quality, radiation dose and amount of contrast agent in group A were evaluated and compared with group B. The dose length product for groups A and B was 62.95±21.54 vs. 160.15±15.13 mGy cm, respectively (t=-23.157, P<0.001). The mean total iodine content was 10.8±0 mg for group A and 22.2±0 mg for group B. In total, 99.4% of the arterial segments could be assessed for the two groups (χ 2=0.267, P=0.606). The results revealed that ultra-low dose one-step high-pitch CTA can provide assessable image quality, and minimize the radiation dose and contrast agent.

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BACKGROUND: Macrophage migration inhibitory factor (MIF) is an inflammatory mediator released by macrophages that is central to the innate immune system, with an upstream role in the inflammatory cascade. MIF is one of the most important pathogenic factors in the development of the autoimmune diseases. In the current study, we investigated the role of MIF in anti-neutrophil cytoplasmic antibody (ANCA)-induced neutrophil activation. METHODS: Plasma levels of MIF from 31 patients with active ANCA-associated vasculitis (AAV) were analyzed by ELISA. The various effects of MIF in ANCA-induced neutrophil respiratory burst and degranulation were measured. RESULTS: Plasma levels of circulating MIF were significantly higher in AAV patients with active disease compared with those in remission and healthy controls. Compared with MIF-primed neutrophils, the MFI value increased significantly in MIF-primed neutrophils further activated with MPO-ANCA-positive IgG or PR3-ANCA-positive IgG (270.8±9.7 vs. 421.5±9.7, P<0.001; 270.8±9.7 vs. 414.1±15.6, P<0.001, respectively). Compared with MIF-primed neutrophils, the lactoferrin concentration increased significantly in the supernatant of MIF-primed neutrophils further activated by MPO-ANCA-positive IgG (567.8±61.2ng/ml vs. 1677.0±42.5ng/ml, P<0.001) or PR3-ANCA-positive IgG (567.8±61.2ng/ml vs. 1546.0±116.2ng/ml, P<0.001), respectively. Interleukin-8 (IL-8), IL-6 and IL-23 were involved in ANCA-induced activation of MIF-primed neutrophils. CONCLUSIONS: MIF primes neutrophils by increasing ANCA antigen translocation. The primed neutrophils can be further induced by ANCA, resulting in respiratory burst and degranulation.

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OBJECTIVE: The goal of this study was to examine the relationship between CCL2-2518A/G, CXCL10-201A/G, and IL8+781C/T gene polymorphism and severity of Enterovirus 71 (EV71) infection in a Chinese population. METHODS: A case-control study was conducted to compare the distribution of genotype and genetic frequency of the CCL2-2518A/G, CXCL10-201A/G, and IL8+781C/T gene polymorphisms among EV71-infected patients (n = 186), including mild cases (n = 103), severe cases (n = 83) and healthy control subjects (n = 233) with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and analyzed the relationship between the CCL2-2518A/G, CXCL10-201A/G, and IL8+781C/T gene polymorphism and the susceptibility to EV71 infection. RESULTS: No significant differences were found in the distribution of genotype CCL2-2518A/G, CXCL10-201A/G, and IL8+781C/T between the healthy control group and EV71-infected patients. However, three SNPs were associated with severity of EV71 infection: the G allele (genotypes AG or GG) in the CCL2-2518A/G (OR 2.34, 95 % CI 1.50-3.65, P < 0.001), the A allele (genotypes AA or AG) in the CXCL10-201A/G (OR 3.60, 95 % CI 1.73-7.47, P < 0.001), and the C allele (genotypes CC or CT) in the IL8+781C/T (OR 2.63, 95 % CI 1.67-4.13, P < 0.001) were more frequent in patients with severe EV71 infection. No significant difference was observed between EV71 encephalitis and severe cases. At the same time, there were significant differences in fever days, WBC, CRP and BG concentration, and CCL2, CXCL10 and IL-8 levels according to the three SNPs among 186 EV71-infected patients, but no significant differences were observed in gender, age, ALT, AST, CK-MB, and CSF evaluations. CONCLUSION: The G carrier of the CCL2-2518A/G, the A carrier of the CXCL10-201A/G, and the C carrier of the IL8+781C/T were found to be associated with severity of EV71 infection, and could be susceptibility factors in the development of EV71 infection in the Chinese population.

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OBJECTIVES: Genetic polymorphism G894T on the endothelial nitric oxide synthase (eNOS) gene has been reported as a susceptibility factor in a number of diseases, but evidence of its effect on enterovirus 71 (EV71) infection is lacking. This study investigated the possible association between this polymorphism (rs1799983) and disease severity in Chinese children with EV71 infection. DESIGN AND METHODS: 185 children with EV71 infection (83 with severe and 102 with mild disease) and 234 control healthy children underwent testing with polymerase chain reaction-restriction fragment length polymorphism (PCR-RLFP) to detect G894T polymorphism. In addition, plasma levels of nitric oxide (NO), interleukin 1 beta (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-α) and serum eNOS activity were measured according to genotype. RESULTS: The presence of GT+TT genotypes and T allele were associated with severe cases compared to genotype GG (OR 2.5, 95% CI 1.2-5.3, P=0.017) and G (OR 2.4, 95% CI 1.2-4.8, P=0.011). Furthermore, in EV71 encephalitis, GT+TT genotype and T allele were also more frequent than GG and G (P<0.05). The NO level and eNOS activity in T carriers (GT+TT) (84.3±2.5µmol/L and 14.4±1.8U/mL) were significantly less compared to in G carriers (GG) (92.0±1.5µmol/L and 19.1±1.7U/mL, P<0.001). But T carriers had higher plasma levels of IL-1ß, IL-6, and TNF-α than people without a T allele (P<0.001), and a significant negative correlation was observed between NO and cytokine levels. CONCLUSION: The results indicate that carrying the T allele of the eNOS G894T gene polymorphism was associated with EV71 infection, and could be a susceptibility factor in the development of EV71 infection in Chinese children.

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Enterovirus 71 (EV71) is one of the common pathogenic agents of hand, foot, and mouth disease (HFMD) and is associated with severe complications including encephalitis. Interleukin (IL)-17F plays an important role in tissue inflammation by inducing release of proinflammatory cytokines and chemokines. We investigated the association between EV71 encephalitis and of IL-17F 7488T/C (rs763780) gene polymorphism, which is known to cause a His-to-Arg substitution at amino acid 161. The study was performed in 58 Chinese patients with EV71 encephalitis and 127 Chinese patients with EV71-related HFMD without complications. Genotyping was determined by the polymerase chain reaction-restriction fragment length polymorphism technique. The patients with EV71 encephalitis had a significantly lower frequency of the IL-17F 7488TC+CC genotypes (10.3%) as compared to the patients with EV71-related HFMD without complications (27.6%, p = 0.008). The frequency of IL-17F 7488C alleles was also significantly lower among the patients with EV71 encephalitis (5.2%) as compared to that of the patients with EV71-related HFMD without complications (15%, OR = 0.310, 95% CI = 0.127-0.756, p = 0.006). Furthermore, homozygotes with the T allele had significantly higher levels of C-reactive protein, white blood cell count, and neutrophil count as compared to the patients with CC+CT genotypes (p = 0.004, 0.001, and 0.000, respectively). These findings suggested that the IL-17F 7488C allele could be significantly associated with protection against encephalitis in Chinese patients with EV71-related HFMD.

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Enterovirus 71 (EV71) often causes large outbreaks of diseases among children worldwide and its pathogenesis remains unclear. The aim of the present study was to investigate the association between interferon-inducible protein 10 (IP-10) polymorphism in children with EV71 infection. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were performed to analyze the gene polymorphisms of IP-10 (-1596C/T) in 58 EV71-infected and 48 control patients. The results showed that in EV71-infected patients the frequency of carrying CT + TT genotype and T allele is 10.3 and 6.0%, respectively, which is significantly lower than that of the controls (29.2 and 15.6%, respectively). Individuals with T allele had a lower risk of EV71 infection [odds ratio (OR) = 0.35, 95% confidence interval (CI), 0.13-0.89]. The results of this study indicated that -1596T allele for the IP-10 gene may be a beneficial factor for EV71 infection.

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Enterovirus 71 (EV71) is one of the common causative agents of hand, foot and mouth disease (HFMD), and is associated with several outbreaks with neurological complications including encephalitis. This study investigated the polymorphisms of interferon gamma (IFN-γ)+874 T/A and interleukin 10 (IL-10)-1082 G/A in 65 Chinese patients with EV71 encephalitis and 113 Chinese HFMD patients without complications. The polymorphisms of IFN-γ+874 T/A and IL-10-1082 G/A were determined by polymerase chain reaction (PCR)-amplification refractory mutation system (ARMS) and PCR-sequence-specific primer (SSP) analysis, respectively. The IFN-γ + 874 A allele was observed with significantly greater frequency in patients with EV71 encephalitis (76.2%) compared with HFMD patients without complications (61.1%, p < 0.01). Similarly, the IL-10 - 1082 A allele was observed with significantly greater frequency in patients with EV71 encephalitis (86.2%) compared with HFMD patients without complications (77.0%, p < 0.05). IFN-γ + 874 A and IL-10 - 1082 A alleles are associated with susceptibility to EV71 encephalitis in Chinese patients.

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