Distinctive Clinical Characteristics and Outcome of ILD-Onset Rheumatoid Arthritis and ACPA-Positive ILD: a Longitudinal Cohort of 282 Cases.

The aim of this study is to investigate the clinical features and outcome of interstitial lung disease (ILD)-onset rheumatoid arthritis (RA) and anti-citrullinated protein antibody (ACPA)-positive ILD-only patients. Arthritis-onset and ILD-onset RA-ILD and ACPA-positive ILD-only patients consecutively admitted to Peking Union Medical College Hospital from January 2008 to December 2017 were enrolled and followed-up. Their demographic, clinical, and laboratory features as well as outcome were collected and analyzed. Compared with arthritis-onset RA-ILD (n = 166, median arthritis-to-ILD interval: 60 months), the ILD-onset RA-ILD (n = 75, median ILD-to-arthritis interval: 2 months) had less rheumatoid nodules and higher titer of ACPA, and manifested more stable ILD (median estimated progression-free survival: 120 vs. 100 months, p = 0.019). Elder age (≥ 65 years) at ILD diagnosis and UIP pattern were associated with ILD progression by both univariate and Cox hazards modeling analysis (p < 0.05). In ACPA-positive ILD-only patients (n = 41), arthritis developed in 7 (17.1%) female patients after a median interval of 24 months. ACPA-positive ILD who subsequently developed arthritis exhibited higher frequency of rheumatoid factor (RF), higher titer of ACPA, and higher levels of ESR and CRP (p < 0.05). Multivariate regression analysis showed that positive RF (OR 12.55, 95% CI 1.31 to 120.48) was the independent risk factor for arthritis development in ACPA-positive ILD-only patients. ILD-onset RA-ILD had more stable ILD compared with arthritis-onset RA-ILD. ACPA-positive ILD patients with positive RF are at increased risk of developing RA.

EZH2 deficiency attenuates Treg differentiation in rheumatoid arthritis.

The chromatin modifier enhancer of zeste homolog 2 (EZH2) methylates lysine 27 of histone H3 (H3K27) and regulates T cell differentiation. However, the potential role of EZH2 in the pathogenesis of rheumatoid arthritis (RA) remains elusive. We analyzed EZH2 expression in PBMC, CD4+ T cells, CD19+ B cell, and CD14+ monocytes from active treatment-naïve RA patients and healthy controls (HC). We also suppressed EZH2 expression using EZH2 inhibitor GSK126 and measured CD4+ T cell differentiation, proliferation and apoptosis. We further examined TGFß-SMAD and RUNX1 signaling pathways in EZH2-suppressed CD4+ T cells. Finally, we explored the regulation mechanism of EZH2 by RA synovial fluid and fibroblast-like synoviocyte (FLS) by neutralizing key proinflammatory cytokines. EZH2 expression is lower in PBMC and CD4+ T cells from RA patients than those from HC. EZH2 inhibition suppressed regulatory T cells (Tregs) differentiation and FOXP3 transcription, and downregulated RUNX1 and upregulated SMAD7 expression in CD4+ T cells. RA synovial fluid and fibroblast-like synoviocytes suppressed EZH2 expression in CD4+ T cells, which was partially neutralized by anti-IL17 antibody. Taken together, EZH2 in CD4+ T cells from RA patients was attenuated, which suppressed FOXP3 transcription through downregulating RUNX1 and upregulating SMAD7 in CD4+ T cells, and ultimately suppressed Tregs differentiation. IL17 in RA synovial fluid might promote downregulation of EZH2 in CD4+ T cells. Defective EZH2 in CD4+ T cells might contribute to Treg deficiency in RA.

Metagenomic profiling of the pro-inflammatory gut microbiota in ankylosing spondylitis.

OBJECTIVE: Gut dysbiosis has been reported implicated in ankylosing spondylitis (AS), a common chronic inflammatory disease mainly affects sacroiliac joints and spine. Utilizing deep sequencing on the feces of untreated AS patients, our study aimed at providing an in-depth understanding of AS gut microbiota. METHODS: We analyzed the fecal metagenome of 85 untreated AS patients and 62 healthy controls by metagenomic shotgun sequencing, and 23 post-treatment feces of those AS patients were collected for comparison. Comparative analyses among different cohorts including AS, rheumatoid arthritis and Behcet's disease were performed to uncover some common signatures related to inflammatory arthritis. Molecular mimicry of a microbial peptide was also demonstrated by ELISpot assay. RESULTS: We identified AS-enriched species including Bacteroides coprophilus, Parabacteroides distasonis, Eubacterium siraeum, Acidaminococcus fermentans and Prevotella copri. Pathway analysis revealed increased oxidative phosphorylation, lipopolysaccharide biosynthesis and glycosaminoglycan degradation in AS gut microbiota. Microbial signatures of AS gut selected by random forest model showed high distinguishing accuracy. Some common signatures related to autoimmunity, such as Bacteroides fragilis and type III secretion system (T3SS), were also found. Finally, in vitro experiments demonstrated an increased amount of IFN-γ producing cells triggered by a bacterial peptide of AS-enriched species, mimicking type II collagen. CONCLUSIONS: These findings collectively indicate that gut microbiota was perturbed in untreated AS patients with diagnostic potential, and some AS-enriched species might be triggers of autoimmunity by molecular mimicry. Additionally, different inflammatory arthritis shared some common microbial signatures.

Changes in lymphocyte subsets in infants with common lower respiratory tract infectious diseases / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the changes and clinical significance of lymphocyte subsets in infants with bronchitis, bronchopneumonia, and bronchiolitis.</p><p><b>METHODS</b>A total of 111 children with bronchitis, 418 children with bronchopneumonia, and 83 children with bronchiolitis were enrolled as disease groups, and 235 healthy children were enrolled as control group. Flow cytometry was applied to measure lymphocyte subsets.</p><p><b>RESULTS</b>The bronchitis group had significantly lower numbers of T cells and CD3+CD8+ T cells than the control group (P<0.05). The bronchopneumonia group had significantly lower numbers of T cells and CD3+CD8+ T cells, a significantly higher number of T helper (Th) cells, and a significantly higher CD4/CD8 ratio than the control group, as well as a significantly higher number of Th cells than the bronchitis group. Compared with the children with mild bronchopneumonia, those with severe bronchopneumonia showed a reduction in T cells and an increase in B cells (P<0.05). The bronchiolitis group had a significantly higher number of Th cells, a significantly higher CD4/CD8 ratio, and a significantly lower number of CD3+CD8+ T cells than the control group (P<0.01). The disease groups showed a significantly higher number of B cells and a significantly lower number of natural killer cells than the control group (P<0.05).</p><p><b>CONCLUSIONS</b>A low, disturbed cellular immune function and a high humoral immune function are involved in the development and progression of lower respiratory tract infectious diseases. The changes in immune function are related to the type and severity of diseases.</p>

Urinary metabolomics and biomarkers of aristolochic acid nephrotoxicity by UPLC-QTOF/HDMS.

BACKGROUND: Drug-induced nephrotoxicity was one of the most important health problems, with increasing morbidity and mortality. Urinary metabolomics based on ultra performance liquid chromatography coupled with quadrupole time-of-flight high-definition mass spectrometry was applied to aristolochic acid (AA) nephrotoxicity rats to characterize the excretion pathways of endogenous metabolites. RESULTS: Compared with the control rats, serum creatinine, serum blood urea nitrogen and urine protein levels were significantly increased in AA nephrotoxicity rats. Metabolomics showed that metabolites including citrate, aconitate, fumarate, glucose, creatinine, p-cresyl sulfate, indoxyl sulfate, hippuric acid, phenylacetylglycine, kynurenic acid, indole-3-carboxylic acid, spermine, uric acid, allantoin, cholic acid and taurine were identified in AA nephrotoxicity rats. CONCLUSION: The identified metabolites suggested that AA nephrotoxicity rats occurred perturbations in Krebs cycle, gut microflora metabolism, amino acid metabolism, purine metabolism and bile acid biosynthesis.

Using the Doubly Charged Selected Ion Coupled with MS/MS Fragments Monitoring (DCSI-MS/MS) Mode for the Identification of Gelatin Species.

In electrospray ionization (ESI) mode, peptides and proteins can be multiply charged ions; in this situation a doubly charged selected ion (DCSI) coupled with mass spectrometry (MS/MS) fragments monitoring (DCSI-MS/MS) method is the most suitable scanning mode to detect known peptides in complex samples when an ion-trap mass spectrometer is the instrument used for the analysis. In this mode, the MS detector is programmed to only select a doubly charged ion as a precursor and to perform continuous MS/MS on one or more of the selected precursors, either during a specific time interval or along the whole chromatographic run. Gelatin is a mixture of high molecular weight polypeptides from the hydrolysis of collagen. In this study, the DCSI-MS/MS monitoring mode was applied to the detection of previously characterized species-specific peptides from different gelatins. The proposed methodology makes use of tryptic digestion for sample preparation and peptide separation and identification by rapid resolution liquid chromatography coupled to an ion trap working in the DCSI-MS/MS mode for the analysis. This methodology was applied to the differential classification of five commercial, homological species of gelatins and proved to be an excellent tool for gelatin product authentication.

High performance liquid chromatography characteristic spectrum of artificial musk.

OBJECTIVE: To determine the high performance liquid chromatography(HPLC)characteristic spectrum of artificial musk. METHODS: HPLC was performed on a HiQ Sil C18 analytical column(4.6 mm×250 mm)with the mobile phase of methanol/water(90:10 v/v);the detection wavelength was 254 nm and the column temperature was maintained at 25 ℃. The similarities among 10 batches of artificial musk samples were anlysised in accordance with the System for Evaluating the Similarities among the Chromatographic Fingerprints of Traditional Chinese Drugs,which is recommended by Chinese Pharmacopoeia Commission. RESULT: The similarities among these 10 batches of artificial musk ranged between 0.999 and 1.000. CONCLUSION: HPLC is a convenient,accurate and reliable technique for establishing the specific spectum of artificial musk and therefore can be used for the quality control of this product.

[Understanding differences between Rheum palmatum and R. franzenbachii from perspective of chemistry, efficacy and toxicity].

Rheum franzenbachii (called Tudahuang in local) has some similarities with R. palmatum (rhubarb) collected by "China Pharmacopoeia" and is often used as a substitute of rhubarb. Can Tudahuang simply replace rhubarb in the application or whether is there difference between Tudahuang and rhubarb, and what is the difference it is important to verify the difference and understand its proper application in the field of clinical practice. In this paper, we discussed the differences of the two herbs from the views of chemistry, efficacy and toxicity based on the author's previous research work as well as literatures, by using the major role of the rhubarb "diarrhea" as the basic point. The analysis result showed that the role of diarrhea Tudahuang was much weaker than that of rhubarb. The reason lies in the difference between the contents of combined anthraquinones component. While acute toxicity in mice of Tudahuang is stronger than that of rhubarb. Thus, Tudahuang should not simply replace rhubarb in practice.

Purgative components in rhubarbs: adrenergic receptor inhibitors linked with glucose carriers.

Rhubarbs and their extractives have been used as cathartic for many years. There have been numerous breakthroughs in the pharmacological research of the drug. However, as the key point of the mechanism, the targets of the effective components still remain unclear. In this paper, with an in vitro system of isolated intestine, we found that both the rhubarb extractives and the anthraquinone derivatives can antagonize the adrenaline effectively. Furthermore, computer based docking provided the binding model of the anthraquinone derivatives and adrenergic receptor. Then, based on the results of the small intestinal promotion and purgative effect experiments in vivo, we built an "inhibitor-carrier" hypothesis to elucidate the mechanism of rhubarb. This work provided key massages for the pharmacological research of rhubarb, such a common and active medicinal plant, and might be of help for the development of new purgative drugs.

Euphorbia kansui roots induced-diarrhea in mice correlates with inflammatory response.

AIM: Euphorbia kansui (E. KS) is a traditional medicine used in China for thousands of years with the effect of propulsion in the gastrointestines. However, there is no reported study of E. KS on gastrointestinal motility until now. The aim of this work is to study the effect of E. KS on the propulsion of gastrointestines, and to elucidate the possible mechanism of action. METHODS: E.KS was prepared as a 30% ethanol extract and used for the experiment of small and large intestines of mice by oral administration with three different dosages (1.2, 0.6 and 0.3 g·kg(-1)). The feces were observed in vivo. The morphology was carried out to detect if there are any changes in the intestines after the extract of E. KS administration. The assays of mRNA and protein expression were employed to observe IL-1ß, TNFα and caspase 3. RESULTS: It was shown that the extract of E.KS promoted diarrhea in mouse feces after administration, inhibited the contraction of smooth muscle of mouse small intestine and caused the inflammatory exudation on the mucosa of the intestines, enhanced the expression of both mRNA and the protein levels of IL-1ß and TNFα in the small or large intestines. CONCLUSION: The results showed that the extract of E. KS acted on the intestinal smooth muscle with propulsion of feces involving the irritation of the intestines with acute inflammatory reactions.

Identification of five gelatins by ultra performance liquid chromatography/time-of-flight mass spectrometry (UPLC/Q-TOF-MS) using principal component analysis.

An ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC/Q-TOF-MS) method coupled with a principal component analysis (PCA) was developed and applied toward identifying donkey-hide gelatin, bovine-hide gelatin, pig-hide gelatin, tortoise shell glue, and deerhorn glue. The UPLC-MS data of the trypsin digested samples were subjected to principal component analysis (PCA) in order to classify these five gelatins. Additionally, marker peptides given by the loadings plot of PCA were identified based on a comparison of recorded LC-MS data with a previously reported database of the corresponding gelatin variants. The results from this study indicate that the proposed method is reliable, and it has been successfully applied to the identification of variants of gelatins commonly used in Traditional Chinese Medicine (TCM).

Serum metabonomics study of adenine-induced chronic renal failure in rats by ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

An ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC Q-TOF MS) metabonomics approach was employed to study the serum metabolic profiling of adenine-induced chronic renal failure (CRF) rats. Acquired data were subjected to principal component analysis (PCA) for differentiating the CRF and the normal control groups. Potential biomarkers were screened by using S-plot and were identified by the accurate mass, isotopic pattern and MS/MS fragments information obtained from UPLC Q-TOF MS analysis. Significant differences in the serum level of creatinine, amino acids and LysoPCs were observed, indicating the perturbations of amino acid metabolism and phospholipid metabolism in adenine-induced CRF rats. This research proved that metabonomics is a promising tool for disease research.

[Systematic study on QAMS method for quality control of Panax notoginseng].

OBJECTIVE: To establish a quantitative method of multi-components by single marker (QAMS) for determining ginsenoside Rg1, Rb1, Rd, Re and notoginsenoside R1 for the purpose of the quality control of Panax notoginseng. METHOD: The relative correction factors (RCFs) between the five active saponins were determined by HPLC-DAD. With any of the five consituents as reference, a QAMS method was established for detect the quantitation of the other four consituents. The durability of the method was evaluated with five different HPLC instruments, five different Cis18 chromatographic columns and four detective wavelengths. Subsequently, the new QAMS method was used to determine the contents of five saponins contained in 43 batches of notoginseng samples, and compare with external standard methods, in order to evaluate the accuracy of the QAMS method. RESULT: When the five saponins were taken for reference, there was no significant difference between the contents of Rg1, Rb1, Rd, Re and R1 contained in the 43 batches of medicines calculated by the QAMS method (Wf) and the content determination result of the external standard method (Ws). The ratio of their results was (Ws/Wr) (94.02 +/- 2.11)%-(99.75 +/- 0.79)%, suggesting that the method was highly accurate. Their relative correction factors showed good durability, ranging between 0.42%-3.7%, 0.52%-3.5% and 0.79%-4.9%, respectively, with different chromatographic columns, different instruments and different detective wavelengths. The relative retention value method could be adopted for accurately position the chromatographic peak of the five consituents, with their values ranging between 0.18%-13%. CONCLUSION: An accurate, rapid and highly durable QAMS method is established for simultaneous determination and location of five saponins, so as to provide reliable basis for the application of the QAMS method in quality control of traditional Chinese medicines.

Selaginellin A and B, two novel natural pigments isolated from Selaginella tamariscina.

Two new unusual natural pigments were first isolated from the whole herbs of Selaginella tamariscina. The structure of selaginellin A (1) was established as (R,S)-4-[(4'-hydroxy-3-((4-hydroxyphenyl)ethynyl)biphenyl-2-yl)(4-hydroxyphenyl)methylene]-2,5-cyclohexadien-1-one and selaginellin B (2) as (R,S)-4-[(4'-methoxy-4-(methyl)-3-((4-methoxyphenyl)ethynyl)biphenyl-2-yl)(4-methoxyphenyl)methylene]-2,5-cyclohexadien-1-one, along with four known biflavonoids, amentoflavone (3), hinokiflavone (4), heveaflavone (5), and 7''-O-methylamentoflavone (6). Their chemical structures were elucidated by spectral analysis of electrospray ionization mass spectroscopy (ESI-MS), one-dimensional nuclear magnetic resonance spectroscopy (1D-NMR) and two-dimensional-nuclear magnetic resonance spectroscopy (2D-NMR) including (1)H-NMR, (13)C-NMR, distortionless enhancement by polarization transfer (DEPT) and heteronuclear multiple bond coherence (HMBC), and single-crystal X-ray diffraction techniques.

A new sesquiterpene isolated from Lindera aggregata (SIMS) KOSTERM.

A new sesquiterpene, neolindenenonelactone (1), was first isolated from the dried roots of Lindera aggregata (SIMS) KOSTERM., along with four known compounds, linderane (2), isolinderalactone (3), linderalactone (4), and 8-hydroxylindestenolide (5). Their chemical structures were elucidated using spectral analysis of fast atom bombardment mass spectroscopy, one-dimensional nuclear magnetic resonance spectroscopy and two-dimensional-nuclear magnetic resonance spectroscopy including (1)H-NMR, (13)C-NMR, distortionless enhancement by polarization transfer, heteronuclear multiple-bond coherence, and single-crystal X-ray diffraction techniques.

Simultaneous determination of ginsenosides in Panax ginseng with different growth ages using high-performance liquid chromatography-mass spectrometry.

The contents of ginsenosides in Panax ginseng not only vary in different parts of the root, but also exhibit yearly variation. In this study, an HPLC-MS method was established in order to simultaneously analyse ginsenosides Rb1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg1 and Rg2. The concentration of ginsenosides in the tap root and root fibre were compared and the yearly variations of nine ginsenosides elucidated. The results indicate that the total content of ginsenosides in the main root and the root fibre both attain a maximum level in the fourth year of growth, although the amount in the former is much higher than in the latter. The variation in the content of ginsenosides during a 2-6 year period suggests that cultivated P. Ginseng can be harvested after the fourth year. The current results will provide useful information for the quality control and good agricultural practice farming of ginseng.

[Study on yield and quality of Scutellaria baicalensis from different habitats].

OBJECTIVE: To compare the economical characters, yield characters and major effective ingredient content of Scutellaria baicalensis from different habitats. METHOD: The characters were studied by field randomized block test combining analysis of variance, correlation analysis and quality analysis. RESULT: There are significant differences in yield and quality of the crude drugs from different habitats. CONCLUSION: The crude drug from Pingyi (Shandong province) is better than those from other habitats.