Enterococcus faecalis Extracellular Vesicles Promote Apical Periodontitis.

Enterococcus faecalis is an important contributor to the persistence of chronic apical periodontitis. However, the mechanism by which E. faecalis infection in the root canals and dentinal tubules affects periapical tissue remains unclear. Bacterial extracellular vesicles (EVs) act as natural carriers of microbe-associated molecular patterns (MAMPs) and have recently attracted considerable attention. In this study, we investigated the role of EVs derived from E. faecalis in the pathogenesis of apical periodontitis. We observed that E. faecalis EVs can induce inflammatory bone destruction in the periapical areas of mice. Double-labeling immunofluorescence indicated that M1 macrophage infiltration was increased by E. faecalis EVs in apical lesions. Moreover, in vitro experiments demonstrated the internalization of E. faecalis EVs into macrophages. Macrophages tended to polarize toward the M1 profile after treatment with E. faecalis EVs. Pattern recognition receptors (PRRs) can recognize MAMPs of bacterial EVs and, in turn, trigger inflammatory responses. Thus, we performed further mechanistic exploration, which showed that E. faecalis EVs considerably increased the expression of NOD2, a cytoplasmic PRR, and that inhibition of NOD2 markedly reduced macrophage M1 polarization induced by E. faecalis EVs. RIPK2 ubiquitination is a major downstream of NOD2. We also observed increased RIPK2 ubiquitination in macrophages treated with E. faecalis EVs, and E. faecalis EV-induced macrophage M1 polarization was notably alleviated by the RIPK2 ubiquitination inhibitor. Our study revealed the potential for EVs to be considered a virulence factor of E. faecalis and found that E. faecalis EVs can promote macrophage M1 polarization via NOD2/RIPK2 signaling. To our knowledge, this is the first report to investigate apical periodontitis development from the perspective of bacterial vesicles and demonstrate the role and mechanism of E. faecalis EVs in macrophage polarization. This study expands our understanding of the pathogenic mechanism of E. faecalis and provides novel insights into the pathogenesis of apical periodontitis.

[Analysis of risk factors for post-polypectomy bleeding and polyp recurrence after colonoscopic polypectomy in children].

Objective: To explore the incidence and the risk factors of post-polypectomy bleeding and polyp recurrence after colonoscopic high-frequency electrocoagulation snare polypectomy. Methods: Clinical data of 1 826 children who underwent colonoscopic high-frequency electrocoagulation snare polypectomy in the Children's Hospital, Zhejiang University School of Medicine from January 2009 to December 2020 was retrospectively analyzed. Demographic characteristics, endoscopic manifestations, pathological features, diagnosis, occurrence of post-polypectomy bleeding and polyp recurrence were collected. The associated risk factors were analyzed by Logistic regression. Results: A total of 1 826 children (1 191 males and 635 females) with 1 967 polypectomies were included. The age was 4.6 (3.2, 6.4) years at initial diagnosis. According to the initial colonoscopy, 1 611 children (88.2%) had solitary polyps, 1 707 children (93.5%) had pedicled polyps, 1 151 children (63.0%) had polyps involving the rectum, and 1 757 children (96.2%) had hamartomatous polyps. Polyposis syndromes were diagnosed in 73 children (4.0%). The post-polypectomy bleeding occurrence was 3.8% (75/1 967). Polyps recurred in 88 children (4.8%). Girls (OR=2.01, 95%CI 1.26-3.23) and sessile polyps (OR=2.28, 95%CI 1.15-4.49) were risk factors for post-polypectomy bleeding (both P<0.05). Multiple polyps (OR=17.49, 95%CI 9.82-31.18), right-colon involvement (OR=3.44, 95%CI 1.89-6.26) and non-hamartoma (OR=2.51, 95%CI 1.04-6.07) were risk factors for polyp recurrence (all P<0.05). Conclusions: Colonoscopic high-frequency electrocoagulation snare polypectomy has low incidence of post-polypectomy bleeding and polyp recurrence. Female patients and sessile polyps have higher risk for post-polypectomy bleeding. Multiple polyps, right-colon involvement and non-hamartoma polyps increase the risk for polyp recurrence.

[Pathological characteristics and clinical prognosis of nodular sclerosis grade 2 of classic Hodgkin's lymphoma].

Objective: To investigate the pathological characteristics and clinical prognosis of nodular sclerosis grade 2 of classic Hodgkin's lymphoma (cHL-NS2) in our cancer center. Methods: A retrospective collection of 23 cases of cHL-NS2 admitted in Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College from July 2008 to April 2019 was performed. Fifty-five cases of nodular sclerosis grade 1 of classical Hodgkin's lymphoma (cHL-NS1) during the same period were selected as control group. Survival curves were plotted using the Kaplan-Meier method, and Cox regression model was used to analyze the influencing factors for survival. Results: The median age of 23 cases of cHL-NS2 was 30 years old. Five cases had extra nodal invasion, and 19 cases were Ⅰ-Ⅱ stage based on Ann Arbor system. The pathological morphology of cHL-NS2 showed that the lymph node structure was completely destroyed and was divided into nodules by thick collagen. The tumor cells in the nodules were abundant and proliferated in sheets. The boundaries between the tumor cells were not clear. The incidence of tumor necrosis in cHL-NS2 was 43.5% (10/23), which was significantly higher than 18.2% (10/55) in cHL-NS1 (P=0.040). The 3-year progression-free survival (PFS) rate of patients in the cHL-NS2 group was 58.1%, which was significantly lower than 89.7% in the cHL-NS1 group (P=0.002). In all of 78 cases, the 3-year PFS rate of patients who did not obtain complete response (CR) was 67.1%, which was significantly lower than 92.2% in patients who achieved CR (P=0.030). Multivariate Cox regression analysis demonstrated that both cHL-NS2 and failure to obtain CR by first-line treatment were independent indicators for short PFS time (P<0.05). Conclusions: In cHL-NS2, the morphology of tumor cells are diverse, and tumor necrosis can be easily found. Under the current first-line treatments of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) or bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone (BEACOPP), cHL-NS2 is an independent indicator for worse PFS.

[Merkel cell carcinoma: a clinicopathological study of 10 cases].

Objective: To investigate the clinicopathological features, differential diagnosis and prognosis of Merkel cell carcinoma (MCC). Methods: The clinical and pathological data of 10 patients with MCC were collected at the 940th Hospital of PLA. The histological characteristics were examined. Immunohistochemical EnVision method was used to detect thyroid transcription factor-1 (TTF1), broad-spectrum cytokeratin (CKpan), CK20, S-100, Ki-67, CD56, chromogranin A, synaptophysin and other markers in the 10 cases. Results: Intradermal MCC of the skin showed a nested, cord-like, cribriform distribution, polygonal cells, uniform size, and lack of cytoplasm. Tumor cell nuclei were large and round, with clear nuclear membranes, fine and scattered chromatin, absence of nucleoli, and mitotic figures of 10 per 50 high power fields. Among them, one patient had sarcoma and squamous cell carcinoma in situ, one patient had squamous cell carcinoma in situ, and one patient had unique cell morphology. Immunohistochemical staining showed that all cancer cells expressed CKpan, synaptophysin and CD56. There were seven cases with perinuclear dot-like positivity of CK20. Six MCCs expressed chromogranin A to varying degrees, while 2 MCCs were weakly positive for p63. The nuclear positive index in the Ki-67 hotspot area was 60%. Conclusion: The histology of MCC varies. Rendering a correct diagnosis of MCC requires adequate sampling, close correlation with clinical history and rational use of immunohistochemical staining. The treatment requires standardized surgery, postoperative radiotherapy and multimodal chemotherapy. Immunotherapy may replace the traditional treatment in the future.

[X-linked inhibitor of apoptosis deficiency manifested as Crohn's disease: a case report and literature review].

Objective: To analyze the clinical characteristics of X-linked inhibitor of apoptosis (XIAP) deficient patients with clinical manifestation of Crohn's disease. Methods: Clinical manifestations, laboratory investigations, genetic testing and therapeutic interventions of one case of XIAP deficiency who was admitted to Department of Gastroenterology in Children's Hospital, Zhejiang University School of Medicine in May 2016 were summarized. PubMed and Chinese database for articles published from January 2016 to June 2017 were searched using the key words of'Crohn's disease'and'XIAP', and the relevant literature was reviewed. Results: The case we reported was a 6-year-1-month-old boy with recurrent bloody stool for 2 months, and abdominal pain with fever for 2 weeks. The patient had a past history of hemophagocytic lymphohistiocytosis (HLH) and epilepsy in the past one year. Complete blood cell count showed mild anemia (Hb108 g/L). The patient had an elevated high-sensitivity C reactive protein (86 mg/L) and erythrocyte sedimentation rate (46 mm/1h) . White blood cells, pus cells and red blood cells were found on routine stool examination. Biochemical panel showed hypoalbuminemia (25.2 g/L) , elevated transaminase (alanine aminotransferase 175 U/L, aspartate transaminase 229 U/L) , hypertriglyceridemia (4.41 mmol/L) , and hyperferritinemia (>1 650.0 µg/L) . Magnetic resonance enterography revealed the intestinal wall thickening and increased enhancement in parts of illeum and colon. Capsule endoscopy revealed multiple ulcers in jejunum. Colonoscopy showed multiple ulcers in colon and the pathological examination revealed chronic inflammation in mucosa of terminal ileum and colon, which was combined with partial necrosis and ulceration. Some phagocytes were seen in bone marrow smears. The patient was given multiple diagnoses, including hemophagocytic lymphohistiocytosis, Crohn's disease, sepsis, epilepsy, severe malnutrition, and hypoproteinemia. The pediatric Crohn's disease activity index (PCDAI) was 37.5. Genetic testing identified a hemizygotic mutation of c.910G>T chrX:123022501 p.G304X in XIAP. The parents had no such mutation. The patient showed response to infliximab with oral intake of mercaptopurine and corticosteroids, and had remission with PCDAI of 0. There were 9 relevant articles (Chinese 0 English 9), which showed 33.3% XIAP deficient patients manifested with inflammatory bowel disease(IBD), who might have other manifestations such as hemophagocytic lymphohistiocytosis or splenomegaly simultaneously or sequentially. Those patients showed poor response to monotherapy. Conclusion: XIAP deficient patients have various clinical manifestations. Genetic testing is important to those male pediatric IBD patients who have the complicated symptoms or little response to standard therapy.

Seroepidemiology of Toxoplasma gondii infection in patients with liver disease in eastern China.

The role of the protozoan parasite Toxoplasma gondii in the pathogenesis of liver disease has recently gained much interest. The aim of this study was to determine the prevalence and risk factors associated with T. gondii infection in patients with liver disease from three cities in Shandong and Henan provinces, China. A case-control study was conducted from December 2014 to November 2015 and included 1142 patients with liver disease and 1142 healthy controls. Serum samples were collected from all individuals and were examined with enzyme-linked immunosorbent assay for the presence of anti-T. gondii IgG and IgM antibodies. Information on the demographics, clinical, and lifestyle characteristics of the participants was collected from the medical records and by the use of a questionnaire. The prevalence of anti-T. gondii IgG was 19·7% in patients with liver disease compared with 12·17% in the controls. Only 13 patients had anti-T. gondii IgM antibodies compared with 12 control individuals (1·14% vs. 1·05%, respectively). The highest seroprevalence was detected in patients with liver cancer (22·13%), followed by hepatitis patients (20·86%), liver cirrhosis patients (20·42%), and steatosis patients (20%). Multivariate logistic regression analysis indicated that consumption of raw meat (odds ratio (OR) = 1·32; 95% confidence interval (CI) 1·01-1·71; P = 0·03) and source of drinking water from wells (OR = 1·56; 95% CI 1·08-2·27; P = 0·01) were independent risk factors for T. gondii infection in liver disease patients. These findings indicate that T. gondii infection is more likely to be present in patients with liver disease. Therefore, efforts should be directed toward health education of populations at high risk of T. gondii infection and measures should be taken to protect vulnerable patients with liver disease.

[Evaluation of the level of urinary cysteinyl leukotriene E4 in diagnosis of bronchopulmonary dysplasia in premature infants].

OBJECTIVE: To explore the correlation of urinary cysteinyl leukotriene E4 (CysLTE4) and diagnosis of bronchopulmonary dysplasia (BPD) in premature infants. METHOD: One hundred and fifty-eight newborn infants were consecutively admitted to the neonatal intensive care units of First Affiliated Hospital of Nanjing Medical University from November 2014 to October 2015.The infants were divided into 3 groups according to the diagnosis on discharge.Sixty-one term infants were classified as having no pulmonary diseases, 52 premature infants were classified as without BPD, and 45 premature infants with BPD were diagnosed at 28 d after birth.Urinary CysLTE4 levels of newborns within 3 days after birth were measured in a blinded way by enzyme- linked immunosorbent assay and were compared among 3 groups, and were evaluated for the diagnostic value and the correlation of gestational age and birth weight.Statistical analysis was performed using correlation analysis, one-way analysis of variance and χ(2) test etc. RESULT: In infants with BPD, the mean urinary CysLTE4 level was (191.0±29.3) ng/L which significantly higher than the premature group without BPD ((164.1±22.7) ng/L) and term infant group ((151.6±41.9) ng/L, F=18.70, P<0.05). Urinary CysLTE4 level within 3 days of life in newborn inversely correlated with gestational age and birth weight (Pearson=-0.33, -0.38, P<0.01). The area under the curve was 0.78, 95%CI: 0.70-0.86, P<0.01, when cutoff was 187.7 ng/L, with Youden index 0.59, sensitivity 77.8% and specificity 81.4%, respectively. CONCLUSION: Urinary CysLTE4 level is up-regulated in BPD infants within early days of life which may be a useful biomarker of early diagnoses of BPD.

MTHFR C677T and A1298C polymorphisms and risk of lung cancer: a comprehensive evaluation.

Results from previous studies on the association between methylenetetrahydrofolate reductase (MTHFR) polymorphisms C677T and A1298C and lung cancer have been conflicting. The aim of this meta-analysis was to clarify the effect of MTHFR polymorphisms on the risk of lung cancer. An electronic search of PubMed, EMBASE, the Cochrane library, and the China Knowledge Resource Integrated Database for papers on C677T and A1298C and susceptibility to lung cancer was performed. The STATA software (Version 13.0) was used for statistical analysis. Statistical heterogeneity, tests of publication bias, and a sensitivity analysis were performed. Twenty-six studies on C677T (12,324 cases and 12,532 controls) and thirteen studies on A1298C (6773 cases and 8207 controls) were included in the meta-analysis. The MTHFR C677T polymorphism showed significant pooled ORs for the homozygote comparison (TT versus CC: OR = 1.518, 95%CI = 1.220-1.890), heterozygote comparison (CT versus CC: OR = 1.053, 95%CI = 0.940-1.179), dominant model (CT + TT versus CC: OR = 1.143, 95%CI = 1.013-1.291), recessive model (TT versus CT + CC: OR = 1.435, 95%CI = 1.190-1.730), and additive model (T versus C: OR = 1.176, 95%CI = 1.066-1.298). In summary, our meta-analysis showed that the MTHFR C677T polymorphism is associated with a significant increase in lung cancer risk in Asian and overall populations, but not in Caucasian populations. However, no significant association between the MTHFR A1298C polymorphism and lung cancer risk was found in either the Caucasian or Asian group with any genetic models.

Performance of HbA(1c) for detecting newly diagnosed diabetes and pre-diabetes in Chinese communities living in Beijing.

AIM: To determine the performance of glycated haemoglobin (HbA(1c)) as a screening tool for detecting newly diagnosed diabetes (NDM) and pre-diabetes. METHODS: A diabetes survey was conducted in Beijing among community dwellers who were willing to participate in the survey. Included in the survey were 903 individuals aged 21-79 years without previously diagnosed diabetes and in whom HbA(1c) and other required covariates had been measured. NDM and pre-diabetes (impaired glucose tolerance + impaired fasting glucose) were defined according to the World Health Organization 1999 criteria based on 75-g oral glucose tolerance test. Receiver operating characteristic curve (ROC) was plotted to determine the performance of HbA(1c). RESULTS: The prevalence of NDM and pre-diabetes was 11.1% and 22.4%, respectively. At an optimal HbA(1c) cut-off point of > or = 6.0%, the test gave a sensitivity of 80.0% and a specificity of 89.8% for diagnosing NDM; at an optimal cut-off point of > or = 5.7%, the sensitivity was 59.4% and specificity 73.9% for diagnosing pre-diabetes. Individuals with HbA(1c)> or = 6.0% tended to be more obese than those with HbA(1c) < 6.0%, but blood pressure and lipid profiles did not differ between the two groups. CONCLUSIONS: HbA(1c) as a single screening test is adequate to detect newly diagnosed diabetes but is not able to identify pre-diabetes in this obese Chinese population.

Risk factors of peripheral arterial disease and relationship between low ankle brachial index and mortality from all-cause and cardiovascular disease in Chinese patients with hypertension.

The aim of this study was to evaluate the risk factors of peripheral arterial disease (PAD) and the relationship between ankle brachial index (ABI) and mortality from all-cause and cardiovascular disease (CVD) in Chinese patients with hypertension. The ABI cohort Study was designed to investigate risk factors of PAD and the relationship between ABI and mortality from all-cause and CVD in Chinese patients. ABI was identified at baseline by measuring systolic pressure at bilateral brachial and tibial arteries. Mortality surveillance was completed from November 2005 to January 2006. Among 3047 participants with hypertension at baseline, 839 (27.5%) were in the low-ABI group. Older age, female gender, higher serum level of triglycerides, lower serum level of high-density lipoprotein, a history of diabetes and a history of smoking were associated with low ABI. During the 13-month follow-up, there were 252 deaths, of which 100 died of CVD. Low ABI was associated with mortality from all-cause and CVD, whose adjusted relative risk was 1.619 (95% confidence interval 1.190-2.203) and 2.454 (1.531-3.933), respectively, in Cox regression models. The survival rate was significantly lower in the low-ABI group than in the normal-ABI group. This study demonstrated that low ABI was independently associated with a high risk of all-cause and CVD mortality in Chinese patients with hypertension. ABI should be promoted as an ideal tool to predict mortality in diabetic patients.