Association of anti-NXP2 antibody with clinical characteristics and outcomes in adult dermatomyositis: results from clinical applications based on a myositis-specific antibody.

OBJECTIVES: The aim was to study and compare the clinical manifestations, auxiliary examinations, and therapeutic responses in patients with different myositis-specific antibody (MSA) types. METHOD: We retrospectively investigated the medical records of 143 hospitalized dermatomyositis patients, all of whom were tested for MSAs, and performed follow-up. Patients were divided into groups with and without anti-nuclear matrix protein 2 (NXP2) antibodies (17 vs 126 patients). Demographic, clinical manifestation (occurring at any time during the disease course), imaging, laboratory, treatment response, and survival data were collected for statistical analyses. RESULTS: Adult dermatomyositis patients with anti-NXP2 antibodies were more prone to dysphagia (P<0.001), had higher levels of muscle injury markers (CK peak, P=0.007; CK peak>1000 IU/L, P<0.001; CK-MB, P=0.002), were younger at onset (P=0.008), and were less likely to present with interstitial lung disease (P=0.016) than the anti-NXP2 antibody-negative subgroup. Multivariable logistic regression analysis showed that onset age (OR=0.96 CI 95%: 0.924-0.999, P=0.043) and dysphagia (OR=7.088, CI 95%: 1.824-27.536, P=0.005) were independent risk factors for anti-NXP2 antibody positivity. Kaplan-Meier survival analysis did not reveal that dermatomyositis patients with anti-NXP2 antibodies have a relatively worse prognosis. However, the disease course was more frequently polycyclic, and 68.75% of patients had a relapsing-remitting disease course. More than half (52.94%) of those who showed no response to treatment used at least 3 disease-modifying antirheumatic drugs. CONCLUSIONS: We show the important clinical features of and risk factors for this unique antibody-mediated form of dermatomyositis. Although these patients had a relatively low mortality rate, they were prone to recurrence, and treatment was challenging. Key points • The clinical features and risk factors for adult dermatomyositis patients with anti-NXP2 antibodies. • The impact of anti-NXP2 antibody on survival outcomes.

Autologous nanofat transplantation accelerates foot wound healing in diabetic rats.

Aim: This study explored the effects of local transplantation of autologous nanofat in the treatment of rats with diabetic foot wounds. Materials & methods: Nanofat was transplanted into the left foot wound of diabetic rats. Phosphate-buffered saline injection in the right served as control. We measured wound size, the extent of epithelization, microvessel density and the expression levels of cytokines at six different time-points postoperation. Results: Compared with the control feet, nanofat-treated feet had significantly smaller wound areas at 7 and 9 days after grafting and showed better re-epithelialization, a greater number of microvessels and higher levels of angiogenic factor expression. Conclusion: This research shows that autologous nanofat transplantation can promote diabetic foot wound healing in rats.

Analysis of the clinicopathological features and prognostic factors of primary hepatic neuroendocrine tumors.

Neuroendocrine tumors (NETs) of the gastrointestinal tract often spread to the liver, while primary hepatic NETs (PHNETs), first described by Edmondson in 1958, are very rare. The majority of existing reports regarding PHNETs have small sample sizes, and the clinicopathological characteristics and prognostic factors are still unclear. The aim of the present study was to analyze the clinicopathological features and explore the prognostic factors of PHNETs. From March 2012 to March 2017, 28 cases of PHNETs were retrospectively evaluated to analyze the clinicopathological features and explore the prognostic factors of PHNETs. The 28 PHNETs patients were males (n=15) and females (n=13) aged between 32 and 76 years (mean=53 years). Among them, 16 patients had clinical symptoms. The remaining 12 patients had no obvious clinical symptoms, only hepatoncus was observed during physical examination. Single-factor analysis showed that carbohydrate antigen 125 (CA125), alanine aminotransferase (ALT), aspartate aminotransferase (AST), hemoglobin (HB), Ki-67 positive index (PI), surgical treatment and pathological grading were correlated to PHNET prognosis (P<0.05); multifactor analysis revealed that Ki-67 PI was associated with the prognosis (P<0.05). Thus, the prognosis of PHNETs may be effectively predicted using the indexes of CA125, ALT, AST, HB, Ki-67 PI, pathological grading and surgical treatment. Pathological classification of grade 3, high expression of Ki-67 PI, abnormal elevation of CA125, abnormalities of ALT and AST, anemia and lack of radical operation indicated a poor prognosis. High expression of Ki-67 PI was an independent prognostic factor for PHNETs.

Expressions of E-cadherin, p120ctn, ß-catenin and NF-κB in ulcerative colitis.

This study was aimed to investigate the expressions of E-cadherin, p120ctn, ß-catenin and NF-κB in ulcerative colitis (UC) tissues and the implications of their expressions in the pathogenesis of UC. The expressions of E-cadherin, p120ctn, ß-catenin and NF-κB were detected by immunohistochemistry, and those of p120ctn and NF-κB by Western blotting in 23 cases of UC and 17 cases of normal colonic tissues. The relationship between the expression of E-cadherin or NF-κB and that of p120ctn was analyzed by Spearman rank correlation analysis. The results showed that in UC and normal colonic groups, the abnormal expression rate of E-cadherin, p120ctn, ß-catenin, and NF-κB was 52.2% vs. 0 (P<0.05), 73.9% vs. 23.5% (P<0.05), 65.2% vs. 17.6% (P<0.05) and 78.4% vs. 23.5% (P<0.05), respectively. p120ctn expression was positively correlated with E-cadherin expression (r=0.404, P<0.05), but negatively with nuclear NF-κB expression (r= - 0.347, P<0.05). Western blotting showed that as compared with the normal controls, the p120ctn protein level was significantly decreased (P<0.05), whereas the NF-κB protein level was increased (P<0.05) in UC tissues. It was concluded that in the colonic tissues of UC patients, the expressions of E-cadherin, p120ctn and ß-catenin are decreased, suggesting the mucosal barrier is impaired in UC. Moreover, NF-κB is increased and activated in the UC tissues, resulting in the inflammation in UC. p120ctn may influence the UC development through modulating intercellular adhesion and inflammatory response.

Pioglitazone Inhibits the Expressions of p22(phox) and p47(phox) in Rat Mesangial Cells In Vitro.

Aim. The purpose of this study was to investigate the effects of pioglitazone on oxidative stress and the expressions of p22(phox) and p47(phox), subunits of NADPH oxidase, in mesangial cells (MCs). Method. Rat mesangial cells were cultured and randomly divided into normal glucose (NG) group, high glucose (HG) group, and pioglitazone group. After 48 h exposure, the supernatants and cells were collected. The expressions of p22(phox) and p47(phox) in MCs were detected by RT-PCR and western blot. The levels of intracellular ROS were determined by flow cytometry. Coloimetry method was used to detect malondialdehyde (MDA) concentrations and superoxide dismutase (SOD) activities. Results. Compared with the NG group, the expression levels of p22(phox), p47(phox) and ROS significantly increased, the activity of SOD decreased in HG group, while the concentration of MDA greatly increased (P < 0.01). Pioglitazone significantly suppressed HG-induced p22(phox) and p47(phox) expressions and oxidative stress. The protein and gene expressions of p22(phox) and p47(phox) were markedly reduced after pioglitazone treatment, so did the ROS generation. The activities of SOD in MCs increased, while the concentrations of MDA in the supernatant decreased greatly by pioglitazone. Conclusions. Pioglitazone can inhibit HG-induced oxidative stress in MCs through suppressing p22(phox) and p47(phox) expressions.

Pioglitazone inhibits the expression of nicotinamide adenine dinucleotide phosphate oxidase and p38 mitogen-activated protein kinase in rat mesangial cells.

BACKGROUND: Oxidative Stress and p38 mitogen-activated protein kinase (p38MAPK) play a vital role in renal fibrosis. Pioglitazone can protect kidney but the underlying mechanisms are less clear. The purpose of this study was to investigate the effect of pioglitazone on oxidative stress and whether the severity of oxidative stress was associated with the phosphorylation level of p38MAPK. METHODS: Rat mesangial cells were cultured and randomly assigned to control group, high glucose group and pioglitazone group. After 48-hour exposure, the supernatants and cells were collected. The protein levels of p22(phox), p47(phox), phosphorylated p38MAPK, total p38MAPK were measured by Western blotting. The gene expressions of p22(phox), p47(phox) were detected by RT-PCR. The levels of intracellular reactive oxygen species (ROS) were determined by flow cytometry. The levels of superoxide dismutase (SOD) and maleic dialdehyde (MDA) in the supernatant were determined respectively. RESULTS: Compared with the control group, the expression levels of p22(phox), p47(phox), phospho-p38 and ROS significantly increased, activity of SOD decreased in high glucose group, while the level of MDA greatly increased (P < 0.01). Pioglitazone significantly suppressed p22(phox), p47(phox) expressions and oxidative stress induced by high glucose. The expressions of p22(phox), p47(phox), phospho-p38MAPK and ROS generation were markedly reduced after pioglitazone treatment (P < 0.05). The activity of SOD in the the supernatant increased (P < 0.05), while the level of MDA decreased greatly by pioglitazone (P < 0.05). The level of oxidative stress was associated with the phosphorylation level of p38MAPK (P < 0.01). CONCLUSION: Pioglitazone can inhibit oxidative stress through suppressing NADPH oxidase expression and p38MAPK phosphorylation.

Development of a solid-phase extraction coupling chemiluminescent enzyme immunoassay for determination of organophosphorus pesticides in environmental water samples.

Solid-phase extraction (SPE) and direct competitive chemiluminescence enzyme immunoassay (dcCL-EIA) were combined for the detection of organophosphorus pesticides (OPs) in environmental water samples. dcCL-EIA based on horseradish peroxidase labeled with a broad-specificity monoclonal antibody against OPs was developed, and the effects of several physicochemical parameters on dcCL-EIA performance were studied. SPE was used for the pretreatment of water samples to remove interfering substances and to concentrate the OP analytes. The coupling of SPE and dcCL-EIA can detect seven OPs (parathion, coumaphos, phoxim, quinalphos, triazophos, dichlofenthion, and azinphos-ethyl) with the limit of quantitation below 0.1 ng/mL. The recoveries of OPs from spiked water samples ranged from 62.5% to 131.7% by SPE-dcCL-EIA and 69.5% to 112.3% by SPE-HPLC-MS/MS. The screening of OP residues in real-world environmental water samples by the developed SPE-dcCL-EIA and their confirmatory analysis using SPE-HPLC-MS/MS demonstrated that the assay is ideally suited as a monitoring method for OP residues prior to chromatographic analysis.

[Epidemiological study of invasive nosocomial candidiasis in 2 teaching hospitals in Beijing].

OBJECTIVE: To investigate the epidemiological characteristics of invasive nosocomial candidiasis in teaching hospitals in Beijing. METHODS: The clinical data of consecutive cases diagnosed as with nosocomial candidiasis hospitalized in Beijing Chaoyang Hospital (CYH) (n=43) and Peking Union Medical College Hospital (PUMCH) (n=34) from January 2004 to September 2006 were retrospectively analyzed to evaluate the incidence and mortality, and the relationship between antifungal therapy and outcomes. The minimum inhibitory concentrations (MICs) for Candida spp were determined by microdilution method. RESULTS: The incidence rates of nosocomial candidiasis of CYH and PUMCH were 0.053% and 0.025% respectively in 2004, and increased to 0.074% and 0.049% respectively in 2006. Candidemia accounted for 55.8% (43/77) of the Candida infection, followed by intra-abdominal infection (26.0%, n=20), intra-thoracic infection (9.1%, n=7), and biliary tract infection (7.8%, n=6). The species mostly frequently isolated were Candida albicans (57.1%), followed by C. tropicalis (19.5%), C. glabrata (14.3%), and C. parasitosis (2.6%). The susceptibility rates of C. albicans, C. tropicalis, C. glabrata, and C. parasitosis to fluconazole were 97.7%, 86.7%, 63.6%, and 100% respectively. No isolates were resistant to amphotericin B. The crude mortality rate of Candida infection was 37.7%. The mortality rate of the candidemia patients who did not receive systemic antifungal therapy was 72.7%, significantly higher than those who received antifungal therapy (37.5%, P = 0.043). Most patients received fluconazole as the first choice (91.7% in PUMCH and 79.2% in CYH) with the survival rates of 63.6% and 52.6% respectively. Conclusion With high morbidity and mortality, invasive nosocomial candidiasis is a big problem in teaching hospitals. Further investigation is necessary to evaluate the incidence and to identify which antifungal agents are most protective for candidiasis.

[Cloning, expression and identification of human survivin in E. coli].

OBJECTIVE: To efficiently express and identify recombinant human survivin in E. coli. METHODS: Survivin cDNA was amplified by reverse transcriptional (RT)-PCR and cloned into the prokaryotic expression vector pBV220, followed by expression of the recombinant plasmid in E.coli strain BL21 (Gold). To obtain survivin protein, DEAE-Sepharose Fast-Flow ion exchange chromatography and Sephacryl S-200 gel filtration were performed. Western blot analysis was used for detecting the expressed product. RESULTS: Survivin protein was expressed in E.coli in the form of inclusion body at the expression level over 30% of the total cell protein. After ion exchange chromatography and gel filtration, the recombinant protein reached a purity over 95% and exhibited specific reaction with mouse anti-human antibody. CONCLUSION: Survivin protein with high purity can be obtained by the method described above to facilitate further study of the anti-apoptosis function of survivin.