[Recommendations on the diagnosis and treatment of anti-neutrophil cytoplasmic antibody associated vasculitis in China].

Anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitis (AAV) is a group of systemic small vasculitis characterized by ANCA positive in serum. Three diseases are included in this group of diseases: granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA) and eosinophilic granulomatosis with polyangiitis (EGPA). In China, standardized diagnosis and treatment of AAV is still lacking. Based on the evidence and guidelines from China and abroad, the Chinese Rheumatology Association formulated the standardization of diagnosis and treatment of ANCA associated vasculitis. The purpose is to standardize the diagnosis of AAV and disease activity assessment, and recommend the treatment strategies.

[High-mobility group protein 1 promotes diethylnitrosamine-induced liver cancer formation in mice by activating mitochondrial biogenesis].

Objective: To study the role of high-mobility group protein 1 (HMGB1) in the promotion of diethylnitrosamine-induced liver cancer formation in C57BL/6 mice and its mechanism. Methods: HMGB1(loxp/loxp)/Alb-Cre(+/-) were used as a liver-specific knockout (KO) of HMGB1 gene in mice. HMGB1(loxp/loxp)/Alb-Cre(-/-), HMGB1(loxp/WT)/Alb-Cre(+/-) and HMGB1(loxp/WT)/Alb-Cre(-/-) born in the same litter were wild-type mice. Six 12-day-old male WT and KO mice were separated and given a single intraperitoneal injection of diethylnitrosamine (25 mg/kg). Six months later, HE staining was used to evaluate the histopathological changes and then the incidence of liver cancer in each mice group was calculated. Serum samples were taken from each mice group to determine alanine aminotransferase levels. Immunohistochemical staining was used to detect the expression and intracellular localizations of HMGB1 protein status in tumor tissue of the two groups of mice. Western blot was used to detect the expressional condition of mitochondrial biogenesis in tumor tissue of the two groups of mice. RT-PCR was used to detect mitochondrial DNA copy number of tumor tissue and normal liver tissue in the two groups of mice. Intra and inter group data comparison was compared using t-tests and one one-way analysis of variance. Results: Compared with WT mice, the liver/body weight ratio of KO mice was decreased significantly (t = 2.634, P = 0.0225). Serum alanine aminotransferase levels in both groups of mice were increased, and the difference was not statistically significant (t = 0.4062, P = 0.6932). There were many visible gray-white nodules of different sizes on the liver surface of WT mice, and the histological type was hepatocellular carcinoma. There was no statistically significant difference in the incidence of liver cancer among different genotypes of WT mice (P > 0.05). The incidence rate of liver cancer in KO mice was significantly reduced (t = 8.521, P < 0.001). Compared with WT mice, the expression levels of HMGB1 and mitochondrial biogenesis (PGC-1α and NRF1) was significantly reduced (t = 6.238, 4.852, P = 0.0335, 0.041) in tumor tissue of KO mice. Mitochondrial DNA copy number was decreased significantly (t = 9.211, P < 0.01). Mitochondrial DNA copy number in tumor tissue of WT mice was significantly higher than that in normal liver tissue (t = 8.305, P = 0.0142). Conclusion: HMGB1 promotes the formation of diethylnitrosamine-induced liver cancer by inducing mitochondrial biogenesis.

Cardiovascular disease in rheumatoid arthritis: medications and risk factors in China.

This study aims to assess the risk factors of cardiovascular disease (CVD) and to determine the association of traditional and biologic disease-modifying anti-rheumatic drugs (DMARDs) with risk for CVD in Chinese rheumatoid arthritis (RA) patients. A cross-sectional cohort of 2013 RA patients from 21 hospitals around China was established. Medical history of CVD was documented. The patients' social background, clinical manifestations, comorbidities, and medications were also collected. Of the 2013 patients, 256 had CVD with an incidence of 12.7%. Compared with non-CVD controls, RA patients with CVD had a significantly advanced age, long-standing median disease duration, more often male and more deformity joints. Patients with CVD also had higher rates of smoking, rheumatoid nodules, interstitial lung disease, and anemia. The prevalence of comorbidities, including hypothyroidism, diabetes mellitus (DM), hypertension, and hyperlipidemia, was also significant higher in the CVD group. In contrast, patients treated with methotrexate, hydroxychloroquine (HCQ), and TNF blockers had lower incidence of CVD. The multivariate analysis showed that the use of HCQ was a protective factor of CVD, while hypertension, hyperlipidemia, and interstitial lung disease were independent risk factors of CVD. Our study shows that the independent risk factors of CVD include hypertension, hyperlipidemia, and interstitial lung disease. HCQ reduces the risk of CVD in patients with RA.

Differential Gene Expression Profiles Reflecting Macrophage Polarization in Aging and Periodontitis Gingival Tissues.

Recent evidence has determined a phenotypic and functional heterogeneity for macrophage populations. This plasticity of macrophage function has been related to specific properties of subsets (M1 and M2) of these cells in inflammation, adaptive immune responses and resolution of tissue destructive processes. This investigation hypothesized that targeted alterations in the distribution of macrophage phenotypes in aged individuals, and with periodontitis would be skewed towards M1 inflammatory macrophages in gingival tissues. The study used a non-human primate model to evaluate gene expression profiles as footprints of macrophage variation in healthy and periodontitis gingival tissues from animals 3-23 years of age and in periodontitis tissues in adult and aged animals. Significant increases in multiple genes reflecting overall increases in macrophage activities were observed in healthy aged tissues, and were significantly increased in periodontitis tissues from both adults and aged animals. Generally, gene expression patterns for M2 macrophages were similar in healthy young, adolescent and adult tissues. However, modest increases were noted in healthy aged tissues, similar to those seen in periodontitis tissues from both age groups. M1 macrophage gene transcription patterns increased significantly over the age range in healthy tissues, with multiple genes (e.g. CCL13, CCL19, CCR7 and TLR4) significantly increased in aged animals. Additionally, gene expression patterns for M1 macrophages were significantly increased in adult health versus periodontitis and aged healthy versus periodontitis. The findings supported a significant increase in macrophages with aging and in periodontitis. The primary increases in both healthy aged tissues and, particularly periodontitis tissues appeared in the M1 phenotype.

Biochip analysis of prostate cancer.

Microarray expression analysis was used to forecast the roles of differentially co-expressed genes (DCG) and DCG and links in the pathogenesis of prostate cancer. In addition, we demonstrate that the relationship between transcriptional factors (TFs) and their targets can be considered a key factor in determining the difference between primary and metastatic prostate cancer. Regulatory impact factors were adopted to calculate the impact of TF. We identified 5 TFs and 29 target genes important in the transition between normal prostate and primary prostate cancer and 2 TFs and 7 target genes important in the transition between primary and metastatic prostate cancer. These results suggest that it may be possible to predict the clinical behavior of prostate cancer based on gene expression analysis.

Screening of feature genes of the renal cell carcinoma with DNA microarray.

AIM: To investigate the underlying molecular mechanisms of renal cell carcinoma (RCC) by using the microarray expression profiles of normal kidney and RCC tissue for early diagnosis and treatment of RCC. MATERIALS AND METHODS: The gene expression profile of GES781 was downloaded from Gene Expression Omnibus database, including including nine tissue samples of RCC tissues removed from nine patients and eight adjacent normal renal tissue samples. We identified the differentially expressed genes (DEGs) by Multtest package in R software. The screened DEGs were further analyzed by bioinformatics methods. Firstly, the comparison of the DEGs expression degree was performed by cluster analysis. Secondly, DAVID was used to perform functional analysis of up- and down- regulated genes and the protein-protein interaction (PPI) networks were constructed by prePPI. Finally, the pathways of genes in PPI networks were discovered by WebGestalt. RESULTS: Compared with the control, we screened 648 down-regulated and 681 up-regulated DEGs. And the down- and up-regulated DEGs with maximum expression degree were UMOD (uromodulin) and FABP7 (fatty acid binding protein 7), respectively. There was significant difference in the gene expression between the normal kidney and RCC tissue. The up-regulated DEGs in RCC tissue were significantly related to the immune responses and the down-regulated DEGs were significantly related to the oxidation reduction. The most significant pathway in the PPI network of UMOD was cytokine-cytokine receptor interaction. CONCLUSIONS: The screened DEGs have the potential to become candidate target molecules to monitor, diagnose and treat the RCC, and might be beneficial for the early diagnosis and medication control of RCC.

Polybacterial challenge effects on cytokine/chemokine production by macrophages and dendritic cells.

OBJECTIVE: To investigate the polymicrobial infection of periodontal disease, which elicits inflammatory mediators/cytokines/chemokines in the local gingival tissues, and a polybacterial challenge of antigen-presenting cells, e.g. macrophages and dendritic cells (DCs), at the mucosal surface. MATERIALS AND METHODS: The cytokine/chemokine profiles of human macrophages and DCs in response to polybacterial challenges were investigated. RESULTS: Oral Gram-negative bacteria elicited significantly greater IL-8 levels from macrophages, compared to Gram-positive bacteria. Gram-positive bacteria did not show synergism in inducing this chemokine from macrophages. In contrast, pairs of oral Gram-negative bacteria elicited synergistic production of IL-8 by macrophages. Similar results were not observed with TNFα, which only appeared additive with the polybacterial challenge. Selected Gram-negative bacterial pairs synergized in IL-6 production by immature DCs. In mature DCs (mDCs), a Porphyromonas gingivalis/Fusobacterium nucleatum and Porphyromonas intermedia/F. nucleatum polybacterial challenge resulted in significant synergism for IL-6 and TNFα levels. However, only the Pi/Fn combination synergized for IL-12 production and there appeared to be no polybacterial effect on IL-10 production by the mDCs. CONCLUSIONS: These results indicate that a polybacterial challenge of cells linking innate and adaptive immune responses results in varied response profiles that are dependent upon the characteristics of the microorganisms that are components of the polybacterial complex.

Supernatants from oral epithelial cells and gingival fibroblasts modulate human immunodeficiency virus type 1 promoter activation induced by periodontopathogens in monocytes/macrophages.

Bacterial and host cell products during coinfections of Human Immunodeficiency Virus type 1-positive (HIV-1(+)) patients regulate HIV-1 recrudescence in latently infected cells (e.g. T cells, monocytes/macrophages), impacting highly active antiretroviral therapy (HAART) failure and progression of acquired immunodeficiency syndrome. A high frequency of oral opportunistic infections (e.g. periodontitis) in HIV-1(+) patients has been demonstrated; however, their potential to impact HIV-1 exacerbation is unclear. We sought to determine the ability of supernatants derived from oral epithelial cells (OKF4) and human gingival fibroblasts (Gin-4) challenged with periodontal pathogens, to modulate the HIV-1 promoter activation in monocytes/macrophages. BF24 monocytes/macrophages transfected with the HIV-1 promoter driving the expression of chloramphenicol acetyltransferase (CAT) were stimulated with Porphyromonas gingivalis, Fusobacterium nucleatum, or Treponema denticola in the presence of supernatants from OKF4 or Gin4 cells either unstimulated or previously pulsed with bacteria. CAT levels were determined by enzyme-linked immunosorbent assay and cytokine production was evaluated by Luminex beadlyte assays. OKF4 and Gin4 supernatants enhanced HIV-1 promoter activation particularly related to F. nucleatum challenge. An additive effect was observed in HIV-1 promoter activation when monocytes/macrophages were simultaneously stimulated with gingival cell supernatants and bacterial extracts. OKF4 cells produced higher levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukins -6 and -8 in response to F. nucleatum and P. gingivalis. Preincubation of OKF4 supernatants with anti-GM-CSF reduced the additive effect in periodontopathogen-induced HIV-1 promoter activation. These results suggest that soluble mediators produced by gingival resident cells in response to periodontopathogens could contribute to HIV-1 promoter activation in monocytes/macrophages, albeit this effect is most notable following direct stimulation of the cells with oral gram-negative bacteria.

Oral bacteria induce a differential activation of human immunodeficiency virus-1 promoter in T cells, macrophages and dendritic cells.

INTRODUCTION: The human immunodeficiency virus (HIV) can integrate into T cells, macrophages and dendritic cells resulting in a latent infection. Reports have also demonstrated that various microbial and host cell factors can trigger HIV reactivation leading to HIV recrudescence, potentially undermining highly active antiretroviral therapies. METHODS: This study evaluated the capacity of oral bacteria associated with chronic periodontal infections to stimulate HIV promoter activation in various cell models of HIV latency. RESULTS: T cells (1G5) challenged with oral bacteria demonstrated a dose-response of HIV promoter activation with a subset of the bacteria, as well as kinetics that were generally similar irrespective of the stimuli. Direct bacterial challenge of the T cells resulted in increased activation of approximately 1.5- to 7-fold over controls. Challenge of macrophages (BF24) indicated different kinetics for individual bacteria and resulted in consistent increases in promoter activation of five fold to six fold over basal levels for all bacteria except Streptococcus mutans. Dendritic cells showed increases in HIV reactivation of 7- to 34-fold specific for individual species of bacteria. CONCLUSION: These results suggested that oral bacteria have the capability to reactivate HIV from latently infected cells, showing a relationship of mature dendritic cells > immature dendritic cells > macrophages > or = T cells. Expression of various pattern recognition receptors on these various cell types may provide insight into the primary receptors/signaling pathways used for reactivation by the bacteria.

Oral infectious diseases: a potential risk factor for HIV virus recrudescence?

As the highly active antiretroviral therapy (HAART) has transitioned human immunodeficiency virus (HIV) infection into a 'chronic disease' management strategy, there is growing evidence that infection with non-HIV pathogens in HIV+ patients may have important public health implications in undermining HAART success and acquired immunodeficiency syndrome progression. Several bacterial and host cell products during infections with non-HIV pathogens have shown the capacity to regulate HIV replication in latently infected cells. A high prevalence of oral infections caused by bacteria, viruses and fungi has been described in HIV+ patients, including periodontal disease. The oral cavity appears to be a site of HIV pathogenesis and potential reservoir for the disease as HIV RNA and DNA forms are present in saliva as well as in gingival crevicular fluid, and oral epithelial cells are susceptible to either cell free or cell-associated HIV infection. The clinical and biological bases of potential associations between chronic oral inflammatory disorders, such as periodontal disease, and exacerbation of HIV viraemia have received little attention. This review attempts to evaluate the current understanding of HIV reactivation as a result of co-infection and/or inflammation induced by non-HIV pathogens in HIV-infected patients, and presents a hypothetic model about the potential role of periodontitis as a global oral infection that potentially contributes to HIV recrudescence.

Improved real-time puff-by-puff GC-MS system for whole smoke analysis.

An automated puff-by-puff mainstream smoke (MSS) system is developed to monitor real-time whole smoke in mainstream cigarette smoke using gas chromatography (GC)-mass spectrometry (MS). The whole-smoke analysis is based on automated sample collection and injection into the GC-MS system. The important feature of this system is the real-time rapid analysis that is simple, sensitive, precise, flexible, and exhibits low carryover of volatile and semivolatile smoke constituents. The system is equipped with an automated sampling and switching valve and a smoking machine. The key improvements of the system, as compared with current and alternative methodologies, include minimizing variations caused by operator sampling techniques, the real-time analysis of MSS, the detection of flavorants in MSS from a single puff of cigarette smoke, the ability to analyze numerous smoke constituents from either whole smoke or the gas phase of a single puff, the ability to monitor a few selected smoke constituents in whole smoke using multiple puffs, and its good feasibility compared with solvent extraction and impinger trapping procedures for volatile organic compounds in MSS. System configuration and sampling methodologies are described. Sensitivity, flexibility, precision, feasibility, carryover, and applications of the system are discussed.

Determination of tobacco alkaloids by gas chromatography with nitrogen-phosphorus detection.

An improved method, gas chromatography (GC) with nitrogen-phosphorus detection (NPD), has been used for determination of alkaloids in green and cured tobacco. Tobacco alkaloids of interest included nicotine, nornicotine, myosmine, anabasine, and anatabine. Tobacco samples were treated with a small quantity of aqueous ammonia solution to "loosen" tobacco tissue and to adjust pH, then extracted with solvent. The composition of the extraction solvent solution affected recoveries of the alkaloids, particularly nornicotine, and also contributed to other phenomena such as carry-over in the injection liner and "quenching" of the nitrogen-phosphorus detector. Use of a packed injection liner (e.g. with Carbowax-KOH on Chromosorb) to reduce carry-over was studied. Quenching of the nitrogen-phosphorus detector was eliminated by reducing the injection volume (i.e. increasing the split ratio), by use of a packed injection liner, and by reducing the amount of pretreatment with aqueous ammonia. A narrow bore capillary column (i.e. 0.18 mm id) was used to improve sensitivity and resolution and to increase the speed of GC analysis. An internal standard, 2,4'-dipyridyl, was used for quantitative measurement of these tobacco alkaloids.

Optimization of headspace sampling using solid-phase microextraction for volatile components in tobacco.

Solid-phase microextraction (SPME) was evaluated as a tool for headspace sampling of tobacco samples. Several experimental parameters (e.g. sampling temperature, pH, moisture, and the type of SPME fibers) were optimized to improve sampling efficiency in two aspects; maximum adsorption and selective adsorption of volatile components onto SPME fibers. The effect of these parameters was often dominated by the physical and chemical nature (e.g. volatility, polarity) of target compounds, thus, SPME sampling conditions can be adjusted to favor a selected group of compounds, such as organic acids in tobacco.

[Genescan analysis of the loss of heterozygosity on the long arm of chromosome 6 in non-small cell lung cancer].

To investigate if there are microsatellite loci in the long arm of chromosome 6 that have close relationship with non-small cell lung cancer, Multiple PCR approach was used to analyze the 36 loci in the long arm of chromosome 6. The PCR products were analyzed in PAGE and then the electrophoresis maps were analyzed with Genescan and Genotyper. There is different LOH frequency in different loci. The total frequency of LOH in 41 lung cancers was 78%(32/41), with the highest frequency of LOH was detected on the locus D6S302(75%). There are 14 loci which have LOH frequency more than 20% and the loci are mainly located in 2 regions: 6q16.3-q21 [6 loci D6S458 (21.43%), D6S1694 (26.92%), D6S1717 (35.71%), D6S1565 (40%), D6S302 (75%), D6S1706 (36.36%) and 6q26-q27 (5 loci D6S1550 (38.46%), D6S264 (20%), D6S1585 (25%), D6S446 (33.33%), D6S281 (30.77%)], There may be tumor suppressor genes located in the two regions, which have a close relationship with non-small cell lung cancer.

Pancreatitis with gallbladder ascariasis in a child: case report.

A 10-year-old girl was admitted for abdominal pain for 1 week. Morning vomiting with 5 Ascaris and diarrhea with Ascaris were found. Radiograph of the abdomen disclosed no significant abnormality. Abdominal sonogram revealed a normal biliary tree; but mildly enlarged pancreatic thickness, and thickened gallbladder wall. Within the thickened gallbladder wall a linear echogenic structure with worm-like movement suspected of being Ascaris was found. We report this case because pediatric pancreatitis and a gallbladder wall thickened with worm-movement have rarely been reported. Urgent treatment and surgery are required for the very ill child with a tensely distended abdomen or signs of peritoneal irritation. Early diagnosis is very important to prevent further complications. We emphasize the role of sonography in the diagnosis of this case and the prevention of progressive deterioration.

Primary spinal epidural non-Hodgkin's lymphoma in a child.

Non-Hodgkin's lymphoma usually involves the central nervous system by metastatic disease. Primary spinal epidural non-Hodgkin's lymphoma (PSENL) is a relatively rare cause of spinal cord compression. It mainly occurs in adults past the 4th or 5th decades. This entity is even rarer in children. The proper treatment modalities are controversial in adults with PSENL. Radiotherapy is the main strategy after surgery; the role of chemotherapy is uncertain. Therapeutic experience in childhood PSENL is extremely limited. We report a 10-year-old boy presenting with backache and bilateral lower leg weakness after minor trauma. Small non-cleaved cell non-Hodgkin's lymphoma of the epidural space was proven after subtotal tumor removal. Other investigations including computed tomography of the chest and abdomen, bone scan, gallium scan, bone marrow aspiration, and cerebrospinal fluid study were all negative for occult disease. The patient received combined therapy with irradiation and chemotherapy after surgery. Esophageal stricture resulting from radiotherapy developed during treatment and colon interposition was performed. He has remained disease free 42 months after the diagnosis with normal functional status.

Bezoars in children: report of four cases.

We reviewed the histories of four children (2 boys and 2 girls, aged 6 mo to 10 yr) with surgically proven bezoars, treated from 1991 to 1998. Two had gastric bezoars and two had intestinal bezoars. All four patients presented with gastrointestinal symptoms and none had undergone previous surgery. Three had a history of abnormal ingestion, including one who ate a pickled fruit from the Boraginaceae (locally known as Po-pu-tsu) plant for 1 year. Plain radiographs revealed intestinal obstruction in all four patients. Abdominal sonography disclosed a hyperechoic band-like lesion and acoustic shadow in both cases of gastric bezoar, and these bezoars were subsequently confirmed by gastric endoscopy. Lower gastrointestinal studies showed obstruction considered most likely to be due to bezoar in one case, while computed tomography (CT) revealed a characteristic small bowel mottled gas pattern in another. Surgical treatment relieved obstruction in all four patients. Bezoar should be suspected in patients with intestinal obstruction who have a history of ingestion of unusual substances. The findings of this report suggest that sonography and endoscopy are useful in the diagnosis of gastric bezoar, while CT is useful in the detection of intestinal bezoar.

Flexible bronchoscopic diagnosis of airway injuries after intubation in children.

BACKGROUND: Airway intubation injuries occur frequently but are often neglected because of spontaneous regression. Although most airway injuries that result from intubation resolve without sequelae, severe complications can develop even when initial symptoms of upper airway obstruction are absent. This retrospective study assessed the clinical features, flexible bronchoscopic findings, and clinical outcomes in children with airway intubation injuries. METHODS: From February 1998 to February 1999, 90 children underwent flexible bronchoscopic examination in our hospital. Fifteen of these patients (6 girls, 9 boys; age range, 1 mo to 5 yr; mean, 21.1 +/- 24.4 mo) were noted to have intubation injuries. RESULTS: The average time for symptoms to emerge after extubation was 1.8 days (range, 0-14 days). The airway intubation injuries diagnosed by flexible bronchoscopy were subglottic stenosis in seven patients, vocal cord granulation in four, supraglottic granulation in two, subglottic web in two, bronchial granulation in two, glottic stenosis in one, and tracheal stenosis in one; two cases were discovered incidentally during postoperative follow-up. Four patients had undergone repeated intubation and eventually required tracheostomy. CONCLUSIONS: The results of this study indicate that flexible bronchoscopy is a simple, safe, and useful technique for the diagnosis and follow-up of airway intubation injury. It should be performed on all patients who have symptoms of upper airway obstruction after extubation as well as those who have undergone long-term or emergency intubation.

Hemorrhagic disease in a newborn due to inadequate vitamin K prophylaxis: case report.

Vitamin K prophylaxis for all neonates has been recommended to prevent hemorrhagic disease of the newborn (HDN), but it is still an uncommon practice in most developing countries throughout the world. In the United States and Canada, where vitamin K injections continue to be recommended in the newborn period, HDN is not a major concern. The risk factors for HDN include inadequate vitamin K prophylaxis, exclusively breast-fed infants, diarrhea, and alternative causes of vitamin K deficiency, such as liver disease and cystic fibrosis. We present an exclusively breast-fed 3-week-old infant with diarrhea for 2 days who died from intracranial hemorrhage related to HDN despite having received a single intramuscular injection of 0.2 mg of vitamin K at birth. Hemorrhage in the infant from vitamin K deficiency should be a concern for pediatricians and obstetricians. We emphasize the importance of administering an adequate dosage of vitamin K for prevention of HDN, particularly in an exclusively breast-fed infant.