肺脏受累的抗中性粒细胞胞浆抗体相关性血管炎患者90例临床特征及预后影响因素分析

目的探讨肺脏受累的抗中性粒细胞胞浆抗体相关性血管炎(AAV)患者的临床特征及预后影响因素。方法将90例肺脏受累的AAV患者的临床资料进行回顾性分析,并随访至2019年9月30日,依据预后将患者分为存活组63例和死亡组27例,比较两组患者的临床资料,采用Kaplan-Meier法进行生存分析,建立Cox回归模型分析肺脏受累的AAV患者预后的因素。结果90例患者中男53例(58.9%),女37例(41.1%),平均年龄为(63.2±12.6)岁。肺脏受累以肺间质性病变多见(52例,57.8%),常合并肾脏损害(61例,67.8%)。死亡组患者发病年龄和诱导缓解期单用糖皮质激素患者比例均高于存活组,淋巴细胞计数、血清白蛋白水平及诱导缓解期使用糖皮质激素联合环磷酰胺患者比例均低于存活组(P<0.05)。Kaplan-Meier生存分析结果显示,年龄≥65岁、淋巴细胞计数<1.1×10^9/L、血清白蛋白<30 g/L、血肌酐≥500μmol/L患者的生存率较低,诱导缓解期接受糖皮质激素联合免疫抑制剂治疗患者的生存率较单用糖皮质激素患者高(P<0.05)。多因素Cox回归分析结果显示,发病时年龄≥65岁、血肌酐≥500μmol/L及诱导缓解期单用糖皮质激素为影响肺脏受累的AAV患者预后不良的独立危险因素(P<0.05)。结论AAV肺脏受累时以肺间质性病变多见,临床表现无特异性,发病时患者年龄≥65岁、血肌酐≥500μmol/L、诱导缓解期单用糖皮质激素为其预后不良的独立危险因素。

慢性鼻窦炎与中性粒细胞的关系研究进展

慢性鼻窦炎(CRS)是一种以上呼吸道局部炎症为特征的异质性疾病。CRS的主要发病机制是各种炎症细胞的浸润,欧洲人种多为嗜酸性细胞浸润的T2型炎症,亚洲人种则多以中性粒细胞为主导的非嗜酸性粒细胞浸润的T1/T3型炎症。中性粒细胞在CRS的整个疾病发生发展过程中都起着举足轻重的作用,中性粒细胞增多可能会影响不同药物对CRS的治疗效果,这类CRS患者对大环内酯类抗生素较为敏感,而对糖皮质激素的治疗反应较差。

Heterogeneity and plasticity of T helper cells

CD4 T helper （Th） cells play critical roles in adaptive immune responses. They recruit and activate other immune cells including B cells, CD8 T cells, macrophages, mast cells, neutrophils, eosinophils and basophils. Based on their functions, their pattern of cytokine secretion and their expression of specific transcription factors, Th cells, differentiated from naive CD4 T cells, are classified into four major lineages, Thl, Th2, Th17 and T regulatory （Treg） cells, although other Th lineages may exist. Subsets of the same lineage may express different effector cytokines, reside at different locations or give rise to cells with different fates, whereas cells from different lineages may secrete common cytokines, such as IL-2, IL-9 and IL-10, resulting in massive heterogeneity of the Th cell population. In addition, the pattern of cytokine secretion may switch from that of one lineage toward another under certain circumstances, suggesting that Th cells are plastic. Tregs are also more heterogeneous and plastic than were originally thought. In this review, we summarize recent reports on heterogeneity and plasticity of Th cells, and discuss potential mechanisms and implications of such features that Th cells display.

Association of the HLA-DRB1*0701 allele with perinuclear anti-neutrophil cytoplasmatic antibodies in Mexican patients with severe ulcerative colitis

AIM： To determine the association between the HLADRB1 alleles and perinuclear anti-neutrophil cytoplasmatic antibodies （p-ANCA） positive in Mexican patients with ulcerative colitis （UC）. METHODS： Ninety Mexican mestizo patients （45 females） with UC, confirmed by biopsy, were studied. High resolution HLA typing was performed by PCR-SSO reverse dot blot and PCR-SSP. Molecular typing techniques were applied to define HLA-DRB1 alleles. Enzyme-linked immunosorbent assay and immunofluorescence techniques were used to detect p-ANCA. RESULTS： Forty-eight （53%） UC patients were positive for p-ANCA by ELISA and IF. We found that p-ANCA- positive UC patients had a significantly increased frequency of HLA-DR7 compared with p-ANCA-negative controls （22% vs 5.1%; pC=0.02, OR=5.2, CI 95%： 1.06-37.82）. Disease activity was scored as severe in 20 patients, moderate in 8, mild in 14 and no activity in the remaining 38 patients according to the Truelove and Witts criteria. Subgroup analysis showed a significantly increased frequency of the HLA-DRB1＊07 allele in 15 of 20 UC patients with severe activity of UC and p-ANCA positivity [100% vs 0%; pC=0.0000001; OR=35]. No significant differences were found between p-ANCA positive patients, HLA-DR alleles and other clinical features such as extraintestinal manifestations, proctocolectomy and extension.CONCLUSION： The HLA-DRB1＊07 is associated with p-ANCA positive UC Mexican patients.

Predictive value of neutrophil infiltration as a marker of Helicobacter pylori infection

AIM： To evaluate the predictive value of neutrophil in- filtration as a marker of Helicobacter pylori （H. pyloni） infection. METHODS： A total of 315 patients with dyspepsia symptoms who underwent upper gastrointestinal en- doscopy were enrolled in this study. Biopsies were evaluated using the updated Sydney system. The medication history of all patients in the preceding 4 wk was recorded. The diagnosis of H. pylori infection was based on 13C-urea breath test at least 4 wk after with- drawal of antisecretory drugs, antibiotics and related drugs. For the patients with subtotal gastrectomy, the diagnosis of H. pylori infection was based on anti-H. pylori immunoglobulin G （IgG） antibody. Serum anti-H. pylori IgG antibody was measured by enzyme-linked immunosorbent assays （Biohit, Finland）. RESULTS： The sensitivity, specificity, positive predic- tive value and negative predictive value of neutrophil infiltration in the diagnosis ofH, pylorlinfection were 92.3%, 83.5%, 77.4% and 94.7%, respectively. Neu- trophil infiltration of gastric mucosa in the histological analysis was strongly associated withH, pylorlinfection （77.4% vs 5.3% in the neutrophil infiltration negative group, P = 0.000）. Moderate neutrophil infiltration was more frequent in H. pylorl infection when compared to mild infiltration （81.8% and 75%, respectively）, but did not reach statistical significance. For those patients with negative rapid urease test, H. pylori was detected in 73.2% of patients with positive neutrophil infiltration on histology. In patients with subtotal gastrectomy, the diagnostic accuracy of neutrophil infiltration in H. pylori infection was 50%. CONCLUSION： Neutrophil infiltration is closely associ- ated withH, pylori and may be recognized as a sign of this infection.

Etiology of inflammatory bowel disease:A unified hypothesis

Inflammatory bowel disease(IBD),including both ulcerative colitis(UC) and Crohn's disease(CD),emerged and dramatically increased for about a century.Despite extensive research,its cause remains regarded as unknown.About a decade ago,a series of findings made me suspect that saccharin may be a key causative factor for IBD,through its inhibition on gut bacteria and the resultant impaired inactivation of digestive proteases and over digestion of the mucus layer and gut barrier(the Bacteria-Protease-Mucus-Barrier hypothesis).It explained many puzzles in IBD such as its emergence and temporal changes in last century.Recently I further found evidence suggesting sucralose may be also linked to IBD through a similar mechanism as saccharin and have contributed to the recent worldwide increase of IBD.This new hypothesis suggests that UC and CD are just two symptoms of the same morbidity,rather than two different diseases.They are both caused by a weakening in gut barrier and only differ in that UC is mainly due to increased infiltration of gut bacteria and the resultant recruitment of neutrophils and formation of crypt abscess,while CD is mainly due to increased infiltration of antigens and particles from gut lumen and the resultant recruitment of macrophages and formation of granulomas.It explained the delayed appearance but accelerated increase of CD over UC and many other phenomena.This paper aims to provide a detailed description of a unified hypothesis regardingthe etiology of IBD,including the cause and mechanism of IBD,as well as the relationship between UC and CD.

Chronic Toxicity of a Novel Recombinant Human Granulocyte Colony-stimulating Factor in Rats

Objective To assess the severity and reversibility of the chronic toxicity of a novel recombinant human granulocyte colony-stimulating factor (rhG-CSFa) in rats and the dose-effect relationship.Methods A total of 100 Sprague-Dawley rats (equal numbers of male and female) were randomly divided into five groups (20 rats in each group):four groups were treated with rhG-CSFa at 500,100,10,1 μg/kg,respectively,and one group was treated with vehicle only to serve as the control.The rats were received subcutaneous injections of rhG-CSFa or vehicle daily for 13 weeks.During the course of the chronic toxicity study,the physical status,body weight,and food consumption were monitored.Half of the rats in each group (n=10) were sacrificed after the last rhG-CSFa administration,and the other half were sacrificed at five weeks after the last rhG-CSFa administration.Urinalyses,blood biochemistry,hematological analysis,histopathological examination,and immunological tests were performed for each of the rats.Results The hematological analyses revealed that the mean white blood cells count,neutrophils count,and neutrophils percentage were increased in male rats at the dose of 10 μg/kg or higher,and these were related with the biological activity of rhG-CSFa.Some small abnormalities were observed in the spleen of a few rats when used highest dose (500 μg/kg,a dosage of 200 folds higher than the normal clinical dosage),but these abnormalities were recovered within 5-week recovery period.No other rhG-CSFa-related abnormalities were observed in this chronic toxicity study.Conclusion No significant toxicity and immunogenicity are observed with rhG-CSFa administration to rats in the chronic toxicity studies.

Clinical Features and Prognosis in 21 Patients with Extrinsic Allergic Alveolitis

Objective To assess the spectrum of causes, clinical features, differences between disease phases, and prognosis of extrinsic allergic alveolitis （EAA）. Methods Patients with EAA diagnosed at Peking Union Medical College Hospital from August 1983 to May 2007 were analyzed retrospectively. Their medical records were examined to gather clinical, laboratorial, radiological, and histopathological data. Patients were divided to three phases （acute, subacute, and chronic） according to clinical presentations. Follow-up data regarding treatment response, subsequent radio- logical and pulmonary function studies, and clinical outcomes were collected. Results A total of 21 cases were enrolled. Among them, 11 were subacute, 10 were chronic. The most common exposure was pet birds （6 cases, 28.6%）. The primary abnormality of pulmonary function was restriction and/or reduction in diffusing capacity （12 cases, 63.2%）. The most common findings on high-resolution computed tomography （HRCT） were ground-glass opacities （13 cases, 68.4%） and centri- lobular nodules （8 cases, 42.1%）. Airway obstruction in pulmonary function test, emphysema, hmg cysts, and fibrosis on HRCT were more frequently seen in chronic than in subacute patients, though the differences were not statistically significant. Bronchoalveolar lavage fluid （BALF） showed lymphocytosis. The total＇cell count and the percentage of neutrophils were significantly higher in subacute than in chronic patients （P〈0.05）. Nonnecrotizing granulomas were seen in 8 （47.1%） cases. Improvement or normalization in symptoms, radiography, and pulmonary function test after treatment were seen in all 18 patients with avail- able follow-up data. Five patients recurred. Conclusions The characteristic abnormalities of pulmonary function, findings on HRCT, and pa- thology are essential for all phases of EAA, and the atypical manifestations such as obstruction and fibrosis can also be present frequently, particularly in chronic cases. Differential cell counts of BALF are related to the phase of the disease. The treatment response and prognosis of EAA are good.

The preoperative neutrophil-to-lymphocyte ratio predicts the outcomes of patients with hepatocellular carcinoma and cirrhosis after hepatectomy

Objective The aim of the study was to investigate the prognostic value of the preoperative peripheral neutrophil-to-lymphocyte ratio （NLR） in patients with hepatocellular cancer （HCC） and cirrhosis after hepa- tectomy. Methods This retrospective study included 321 patients with HCC who underwent resection. The NLR was calculated using the neutrophil and lymphocyte counts in routine preoperative blood tests. Receiver operating characteristic curve analysis was performed to select the most appropriate NLR cutoff value. The preoperative NLR, patient demographics, and clinical and pathological data, including disease-free survival （DFS） and overall survival （OS）, were analyzed. Results The NLR was correlated with alpha-fetoprotein levels （X2 = 5.876, P = 0.015）, tumor size （X2 = 32.046, P 〈 0.001）, portal vein tumor thrombus （PVTT; x2 = 4.930, P = 0.026）, tumor encapsulation （x2 = 7.243, P = 0.007）, and recurrence （x2 = 7.717, P = 0.005）. Multivariate analyses illustrated that the number of tumors, PVTT, tumor size, and the NLR were independent factors for predicting DFS and OS. in patients with HCC and cirrhosis, but not among those without cirrhosis, a larger NLR predicted poorer postoperative DFS and OS （both P 〈 0.001）. Conclusion As a simple, effective independent predictor for patients with HCC, the preoperative NLR plays an important role in accurately predicting the postoperative outcomes of patients with HCC and cir- rhosis, but not those of patients without cirrhosis.

Changes in the number of circulating CD34＋ cells after eccentric exercise of the elbow flexors in relation to muscle damage

Background： It has been reported that strenuous exercise increases the number of bone marrow-derived progenitor cells such as CD34＋ cells in the blood, but no previous studies have investigated the changes in circulating CD34＋ cells following resistance exercise. This study tested the hypothesis that the number of CD34＋ cells in the blood would increase after eccentric exercise of the elbow flexors, but decrease in recovery, and the magnitude of the changes would be dependent on the magnitude of muscle damage. Methods： Nine men （28.0 ± 6.6 years） performed exercises consisting of 10 sets of six maximal voluntary eccentric contractions of the elbow flexors with their non-dominant arm. Six of them performed the same exercise with the same ann 4 weeks later. Changes in indirect markers of muscle damage were measured before, within 10 min after, and at 24, 48, 72, and 96 h after eccentric exercise. Differential leukocyte counts （total leukocytes, neutrophils, lymphocytes, monocytes） and CD34＋ cells in the blood were measured before, immediately after, and at 2, 24, 48, 72, and 96 h following the exercises. Results： After eccentric exercise, significant （p 〈 0.05） decreases in maximal voluntary isometric contraction torque and increases in delayed onset muscle soreness and plasma creatine kinase activity were observed. However, no significant changes in leukocytes and CD34＋ cells were evident. The changes in muscle damage markers were significantly （p 〈 0.05） smaller following the second exercise session as compared with the first exercise session, but the changes in leukocytes and CD34＋ cells were not significantly different between sessions. Conclusion： These results did not support the hypothesis, and showed that eccentric exercise-induced muscle damage to the elbow flexors did not influence the number of circulating CD34＋ cells.

Influence of Photoperiod over Morphometric and Hematological Parameters of Juvenile Piracanjubas （Brycon orbygnianus）

The aim of this study was to evaluate morphometric and hematological parameters of juveniles Piracanjubas treated with two distinct photoperiod. Twenty nine fish with 100 g, in a factorial design were performed in 18 h light：6 h dark （18L：6D） and 6L：lSD two different photoperiod, and morphometric and hematological parameters were evaluated at 45 d and 90 d after photoperiod treatment. Benzocaine 1% was used for euthanasia. Fish were weighed and measured, so as visceral fat, liver and gonads. Hematocrit, total count of white cells, lymphocytes and neutrophils were counted. Results showed that there was no significant relationship between bodyweight and standard length. Significant interaction （P = 0.00018） had been found between the fat index （FI） and gonado-somatic index （GSI） and the photoperiod and time. At 90 d, the fish kept in 18L：6D had a lower rate of fat than the fish kept in 6L：18D （P 〈 0.05）. The GSI was higher in the group 6L.＇18D compared with the 18L：6D （P 〈 0.005）. There was no interaction between photoperiod and period of treatment for total leukocyte count, however with 90 d, the total leukocyte count was decreased （P 〈 0.05）. Moreover, decrease of the hematocrit from 58.1% to 46.9% and an increase of lymphocytes from 31.5% to 43.5% had been found in fish kept in 18L：6D after 90 d （P 〈 0.05）. Fish kept in 6L：ISD presented an increase in segmented neutrophils and lymphocytes （P 〈 0.05）. It is concluded that photoperiod maintained at 6L： 18D for Piracanjubas permits a greater gonadal development and an amount of immune cells over the photoperiod maintained in 18L： 6D.

Expression of HNPlcDNA in CHO-dhfr^- cells

Objective: To prepare secretary recombinant human neutrophil peptidel (HNP1)and test its antimicrobial activity. Methods: The eukaryotic expression vector pcDNA3. 1/V5-His-TOPO-HNP1 was cotransfected with plasmid pDCH1P11 carrying dhfr gene into dhfr- negative CHO (CHO-dhfr- ) cells and recombinant protein was verified by ELISA; G418 selective medium was used to screen the stably expressing cell clones followed by serial passages in 5×10-8 mol/L and 5×10 mol/L methotrexate (MTX) for gene amplification. Finally 4 cell clones with high expression level were obtained and confirmed by ELISA, RT-PCR and IFA. The bacteriastatic activity of concentrated supernatants was tested in vitro as well. Results: The expression level of recombinant HNPl ranged from 18.85 mg/L·48 h to 47.46 mg/L·48 h per 106 cells that was almost 200-fold increase than that in G418 selective medium. 303 bp segments were amplified from 4 stably tranfec tant clones which matched the length of HNPl cDNA by RT-PCR. Strong fluorescence was visible in cell plasma in the sta blly transfectant cells by IFA. K-B disc agar diffusion test showed obvious bacteriastatic diffusion on MH plate of E. coli. Conclusion: HNP1cDNA can be strongly expressed in CHO-dhfr- cells, which supernatants exhibited high inhibitive effect against bacteria.

Staining Effect of Pomegranate Flower Extract on Human Blood Cells： First Results

Pomegranate flower plant, popularly known using for the treatment of various diseases, was not investigated as a source of dye for cytological studies using human blood cells. The importance of this study is to appear dyeing result of pomegranate flower extract on human blood cells. The natural dye source was pomegranate flower known as roselle and potassium aluminum sulfate （alum = KAISO4＇ 12H20） was used as mordant or metal salt. Distilled water was used as solvent. Fresh, clean and air-dried flowers were extracted with distilled water at 100℃ for 30 minutes and then filtered. One drop blood from a healthy 20-year woman was spread as a peripheral on to ten plates and dried at nearly 25℃. These slides were stained by soaking in pomegranate flower extract with/without alum （KA1SO4·12H2O） at 100℃ for 60 minutes. Slides were washed with distilled water, dried and done microscobic examination. The different blood cells dyed dark orange in alum mordant media at 100 ℃. As a result, pomegranate flower has the capacity to use dyeing human blood cells such as eosinophil, basophil and neutrophil.

Expression characteristics of neutrophil and mononuclear-phagocyte related genes mRNA in the stable angina pectoris and acute myocardial infarction stages of coronary artery disease

Objective To investigate expression differences of neutrophil and mononuclear phagocyte related gene mRNAs among acute myocardial infarction （AMI）, stable angina （SA） and control groups, and then discuss their expression characteristics in the stable angina pectoris （SAP） and AMI stages of coronary artery disease （CAD）. Methods Whole Human Genome Oligo Microarrays were applied to assess the differential expression characteristics of neutrophil and mononuclear phagocyte related mRNAs in patients with AMI （n = 20）, SA （n = 20） and controls （n = 20）. Results （1） Almost all colony-stimulating factors （CSF） and their receptors related mRNAs was up-regulated in AMI and SA groups compared with the control group, and the expression of granulocyte-macrophage colony stimulating factor receptor （GM-CSFR） and granulocyte colony stimulating factor receptor （G-CSFR） mRNAs in the AMI group was significantly up-regulated compared with the other two groups （P 〈 0.01）. （2） The expression of mRNAs related to monocyte chemoattractant protein-1 （MCP-1）, CCR2 （MCP-1 receptor） and CXCR2 （IL-8 receptor） was significantly up-regulated （P 〈 0.01） in AMI group compared with SA and control groups IL-8 mRNA expression in the AMI group was clearly higher than the controls （P 〈 0.05）. （3） All mRNAs expression related to opsonic re- ceptors （IgG FoR and C3bR/C4bR） was significantly up-regulated in AMI group compared with SA and control group （P 〈 0.01）, and the SA group showed an upward trend compared with controls. （4） Most pattern recognition receptor （PRR）-related mRNAs expression was up-regulated in AMI group compared with SA and control groups. Most toll-like receptor （TLR） mRNAs expression was significantly up-regulated （P 〈 0.01） than the SA and control groups, macrophage scavenger receptor （MSR） mRNA was significantly up-regulated in AMI group compared with the control group （P 〈 0.01）, and the SA group showed an upward trend compared with the controls. Conclusions The expression of most neutrophil and mononuclear-macrophage function related genes mRNAs was significantly up-regulated by stages during the progression of CAD, suggesting that the adhesive, chemotactic and phagocytic functions of neutrophil and mononudear-macrophage were strengthened in the occurrence and development of coronary atherosclerosis and AMI. This also showed a stepped up- ward trend as the disease progressed.

ANTI-INFLAMMATORY EFFECTS OF MAGNOLIAE FARGESII VOLATILE OIL

Objective To explore the anti-inflammatory effects of magnoliae fargesii volatile oil.Methods Human umbilical vein endothelial cells (HUVECs) were stimulated by TNF-α to express the adhesion molecules. Then the anti-adhesion effects of magnoliae fargesii volatile oil between HUVECs and human peripheral neutrophils were observed. The ischemia-reperfusion animal models were established by 60min renal ischemia followed by 1, 3, 6 and 24h reperfusion. Rats were randomly divided into the following groups: the sham-operation controls, ischemic group only treated with normal saline, and treated group infused magnoliae fargesii volatile oil before reperfusion. Then the renal injury of rats was detected. Results High rate of cell adhesion between HUVECs and neutrophils was observed. Magnoliae fargesii volatile oil could inhibit the adhesion process at the concentration of 0.5μL/mL (191.6±8.6), 1.0μL/mL (158.2±9.0) and 2.0μL/mL (155.2±9.7) (P<0.05). The anti-adhesion effects were strengthened with the increase of volatile oil concentration. Blood urea nitrogen and creatinine levels of the animal models were significantly increased after 24h reperfusion while the increase was remarkably attenuated by the treatment with magnoliae fargesii volatile oil. The renal injury was severe after 1h reperfusion, which was significantly attenuated by the treatment of magnoliae fargesii volatile oil. Conclusion Magnoliae fargesii volatile oil has anti-inflammatory effects.

Polysaccharide sulfate 916 inhibits neutrophil-endothelial adhesion

OBJECTIVE: To study the effect of polysaccharide sulfate 916 (PS916) on neutrophil-endothelial cell adhesion. METHODS: Cell adhesion was evaluated by testing neutrophil myeloperoxidase activity. Expression of adhesion molecule in human umbilical vein endothelial cell (HUVEC) was measured by ELISA. The neutrophil activation rate induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) was tested by nitroblue tetrazolium (NBT) reduction. RESULTS: Tumor necrosis factor alpha (TNFalpha, 50 - 800 U/ml) increased the adherence of neutrophil to TNFalpha-stimulated HUVEC in a concentration and time dependent manner. PS916 (0.01 - 1.0 mg/ml) dose-dependently inhibited the adherence of neutrophils to TNFalpha-stimulated HUVEC. fMLP increased the activation rate of neutrophils independent of concentration. PS916 also inhibited the adherence of fMLP-activated neutrophils to HUVEC. Moreover, PS916 inhibited adhesion molecule expression in TNFalpha-stimulated HUVEC. CONCLUSIONS: PS916 inhibited neutrophil-endothelial adhesion. The mechanism of its action was partially related to suppressing the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1).

A simple method for removing low-density granulocytes to purify T lymphocytes from peripheral blood mononuclear cells

Objective： Low-density granulocytes （LDGs） can form neutrophil extracellular traps （NETs） spontaneously and excessively. When peripheral blood mononuclear calls （PBMCs） are used for studying T lymphocytes, LDGs contained in the PBMCs may decrease the threshold of activating T lymphocytes by forming NETs. This study focused on the profiles of LDGs in common autoimmune diseases and methods for removing LDGs from PBMCs. Methods： The percentages of LDGs in PBMCs from 55 patients with dermatomyositis （DM）, 15 with polymyositis （PM）, 42 with rheumatoid arthritis （RA）, 25 with systemic lupus erythematosus （SLE）, and 19 healthy controls were determined by flow cytometry. Three methods of removing LDGs were explored and compared. After removal, PBMCs from six patients with positive T-SPOT.TB were tested again to find out if LDGs contained in the PBMCs could influence T lymphocyte reactions. Results： Significantly higher LDG percentages were found in PBMCs from patients with DM （（8.41±10.87）%, P〈0.0001 ）, PM （（8.41±10.39）%, P〈0.0001 ）, RA （（4.05±6.97）%, P=0.0249）, and SLE （（7.53±11.52）%, P=0.0006）, compared with the controls （（1.28±0.73）%）. The T-SPOT.TB values significantly decreased after LDGs were removed. Increasing relative centrifugal force （RCF） within a limited range can decrease the LDG percentage from an initial high level, but not markedly increase the LDG clearance rate. Compared with the whole blood sediment method, the PBMC adherence method can significantly remove LDGs yet scarcely influence the T lymphocyte percentage in PBMCs. Conclusions： The LDG percentage in PBMCs is significantly increased in patients with S/E, DM, PM, and RA. The influence of LDGs on T lymphocytes cannot be ignored in PBMC cultures. The adherence method is a simple and easy-to-use method for removing LDGs and purifying T lymphocytes from PBMCs.

Metadoxine inhibits the infiltration of macrophages and neutrophils into liver tissue in acute alcoholic liver injury

Alcohol consumption causes significant liver damage,including hepatitis,fibrosis,cirrhosis,and even primary liver carcinoma.Metadoxine(MTDX)is considered to be a beneficial treatment for alcoholic liver disease(ALD)because it accelerates the metabolism and elimination of ethanol.However,the underlying mechanism is not well understood.Here,the rat model of ALD was developed by feeding with 50%ethanol at the dose of 5 g/kg,and samples of serum and liver tissue were collected to test the levels of liver injury and inflammation and evaluate the hepatoprotective function of MTDX in alcohol-induced liver injury.Further investigation on the infiltration of immune cells was performed to understand the potential hepatoprotective mechanism of MTDX in the ALD model.The results showed that MTDX attenuated liver injury,evidenced by decreased levels of alanine transaminase(ALT),aspartate aminotransferase(AST),and alkaline phosphatase(ALP).Meanwhile,the liver proinflammatory environment was improved after MTDX treatment,evidenced by decreased levels of TNF-α,IL-6,and NLRP3 in the liver tissue.Furthermore,inhibited infiltrations of macrophages and neutrophils were observed in MTDX-treated ALD rats compared with the untreated ALD rats.Our results indicated that MTDX played an important role in preventing the progression of ALD,and the underlying mechanisms might be related to its function of attenuating liver inflammation by inhibiting immune cell infiltration.

Dynamic process of phagocytosis and forms of macrophage cell death induced by ingestion of apoptotic neutrophils

Clearance of apoptotic neutrophils by macrophages is important for both the successful resolution of acute inflammation and homeostasis.However,the dynamic process of phagocytosis of apoptotic neutrophils by macrophages and the fate of macrophages after the ingestion of apoptotic neutrophils has not been well documented.In the present study,we staged the recognition and tethering,internalization,digestion and exocytosis steps of phagocytosis of apoptotic neutrophils.Furthermore,we found that after the ingestion of apoptotic cells,a subset of macrophages underwent cell death by autophagy,apoptosis or oncosis as revealed by transmission electron microscopy and confocal microscopy combined with specific dyes.The percentage of autophagic,apoptotic and oncotic macrophages were 8.00%±2.00%,12.33%±2.08%,and 3.66%±1.50%,respectively.These results indicated that after ingestion of apoptotic neutrophils,a subset of macrophages undergoes autophagy and apoptosis.We propose that autophagy of macrophages after the ingestion of apoptotic cells may be a new mechanism present in the resolution of inflammation.

Growth model of a hydrothermal low-temperature Si-rich chimney:Example from the CDE hydrothermal field,Lau Basin

The CDE hydrothermal field was first discovered during a Chinese cruise to the East Lau Basin Spreading Centre in 2007. Apart from significant amounts of loose Fe-Si-Mn （oxyhydr） oxide （referred to as oxide below） precipitates, a small Si-rich oxide chimney was also recovered on this cruise. In this study, we report on the mineralogical and geochemical analyses of this chimney and a model for its growth that has been developed. Based on the mineralogy and O isotope results, the chimney walls can be divided into four growth generations （layers） from the inner to the outer layers： amorphous opal and barite layer （pre- cipitation temperature 68.5℃ based on oxygen isotope determinations）, a rod-like amorphous layer （precipitation temperature 39.6℃）, a filamentous Fe-Si oxide layer, and an outer Fe-Mn oxide layer. Investigations based on SEM and EDS showed that neutrophilic Fe-oxidizing bacteria play an important role in the formation of this chimney, particularly in the outer two genera- tions. In the first stage, the metabolic activity of the microbes results in the pervasive precipitation of the filamentous Fe-rich oxides inside a ring formed by some amorphous opal and barite; therefore, a loose porous layer forms. In the second stage, amorphous opal then precipitates inside this wall as a result of conductive cooling and gradually controls the mixing between the hydrothermal fluids and ambient seawaters. In the third stage, barite and some amorphous opal form from the higher tem- perature fluids at the summit of the chimney growth history. In the last stage, the chimney wall becomes thicker and denser and the exchange of hydrothermal fluids and seawater ceases. As a result, a Fe-Mn oxide layer precipitates onto the outer surface of the chimney wall as neutrophilic Fe-oxidizing bacteria reoccupy the surface of the chimney. This mineral sequence and the resultant growth generations are confirmed by the chemical characteristics of the chimney wall. Sr isotopes extracted from the Fe oxides of the four-generation wall generally show a decreasing trend of the 87Sr/86Sr ratios from the second layer to the in- ner layer （from 0.707008 to 0.705877） except for the outer layer （0.706502）. The Sr isotope and chondrite normalized REE patterns of the corresponding bulk samples from the chimney wall also display a similar trend. Our study shows that the bio- genic filament network plays a key role in the formation of the chimney in contrast to previous growth models of higher temperature chimneys, which often ignore the influence of biogenic factors.