Effects of neutrophil DNA methylation on periodontitis and research progress in bioinformatics / 国际生物医学工程杂志

Periodontitis is a chronic infectious disease leading to periodontal connective tissue destruction and alveolar bone resorption, which is widely prevalent and seriously endangers the oral and systemic health of a wide range of patients. The host immune inflammatory response plays a major role in the tissue destruction of periodontitis. Polymorphonuclear neutrophils (PMNs), as one of the important immune cell components in periodontal tissues, can trigger the host immune inflammatory response through the release of pro-inflammatory factors, which in turn leads to periodontitis. DNA methylation can influence the function of immune cells by regulating gene expression. Bioinformatics technology can provide new ideas for the treatment of periodontitis by analyzing the gene expression profiles and DNA methylation data of periodontal tissues from public databases of periodontitis patients and healthy populations, uncovering key DNA methylation genes of PMNs, and elucidating the influence of these genes on the pathological progression of periodontitis.

Experimental study of transplantation of human umbilical cord mesenchymal stem cells in the treatment of alkali-burned cornea / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the efficacy of transplantation of human umbilical cord mesenchymal stem cells(hUCMSCs)in the treatment of corneal alkali burn in rabbits, and study the infiltration of polymorphonuclear neutrophils(PMNs)and the changes of vascular endothelial growth factor(VEGF)expression.METHODS: Corneal alkali burn models were established in right eyes of 75 healthy Japanese white rabbits, which were divided into three groups(group A, B and C), with 25 rabbits in each group. Group A was treated with amniotic membrane combined with hUCMSCs on the day after corneal alkali burn. Group B was treated with amniotic membrane only. Group C did not give any treatment after corneal alkali burn. At 3, 7, 14, 21 and 28d after corneal alkali burn, the corneal recovery was observed by slit lamp and photographed, the growth of corneal neovascularization(CNV)was scored, and corneal tissue was separated to make pathological sections. PMNs infiltration was observed by hematoxylin-eosin(HE)staining, and the expression of VEGF was determined by immunohistochemical staining.RESULTS: The growth of CNV in group A was much slower than that in group B at 14d after alkali burn. The CNV growth score around lesions of group A was significantly lower than that of group B(P<0.05). The quantity of PMNs increased on the 3d with the stromal layer of cornea infiltrated, relatively decreased on the 7d, shown a peak on the 14d, and then decreased gradually. Early infiltration after alkali burn was in the corneal stroma of the lesion area, and the extent of infiltration was equal to the ulcer area at later stage. The cell densities of corneal PMNs in group A and group B were significantly lower than those in group C at all time points after alkali burns(P<0.05), and those in group A were significantly lower than group B at 14 and 21d(P<0.05). The expression levels of corneal VEGF in all groups after alkali burn reached peak at 7～14d and decreased significantly at 28d, and the expression levels of VEGF in group A and group B at all time points after alkali burn were significantly lower than those in group C(P<0.05), and group A was significantly lower than that in group B at 7, 14 and 21d(P<0.05).CONCLUSION: The transplantation of hUCMSCs after alkali burn cornea can reduce the formation of CNV and inhibit corneal revascularization after alkali burn. The corneal pathological lesions and vascularization are closely related to PMNs and VEGF.

Inertial label-free sorting and chemotaxis of polymorphonuclear neutrophil in sepsis patients based on microfluidic technology / 生物医学工程学杂志

Reduced chemotactic migration of polymorphonuclear neutrophil (PMN) in sepsis patients leads to decreased bacterial clearance and accelerates the progression of sepsis disease. Quantification of PMN chemotaxis in sepsis patients can help characterize the immune health of sepsis patients. Microfluidic microarrays have been widely used for cell chemotaxis analysis because of the advantages of low reagent consumption, near-physiological environment, and visualization of the migration process. Currently, the study of PMN chemotaxis using microfluidic chips is mainly limited by the cumbersome cell separation operation and low throughput of microfluidic chips. In this paper, we first designed an inertial cell sorting chip to achieve label-free separation of the two major cell types by using the basic principle that leukocytes (mainly granulocytes, lymphocytes and monocytes) and erythrocytes move to different positions of the spiral microchannel when they move in the spiral microchannel under different strength of inertial force and Dean's resistance. Subsequently, in this paper, we designed a multi-channel cell migration chip and constructed a microfluidic PMN inertial label-free sorting and chemotaxis analysis platform. The inertial cell sorting chip separates leukocyte populations and then injects them into the multi-channel cell migration chip, which can complete the chemotaxis test of PMN to chemotactic peptide (fMLP) within 15 min. The remaining cells, such as monocytes with slow motility and lymphocytes that require pre-activation with proliferative culture, do not undergo significant chemotactic migration. The test results of sepsis patients ( n=6) and healthy volunteers ( n=3) recruited in this study showed that the chemotaxis index (CI) and migration velocity ( v) of PMN from sepsis patients were significantly weaker than those from healthy volunteers. In conclusion, the microfluidic PMN inertial label-free sorting and chemotaxis analysis platform constructed in this paper can be used as a new tool for cell label-free sorting and migration studies.

Neutrophils activated by BJcuL, a C-type lectin isolated from Bothrops jararacussu venom, decrease the invasion potential of neuroblastoma SK-N-SH cells in vitro

ABSTRACT Background: Neuroblastoma is a pediatric tumor with a mortality rate of 40% in the most aggressive cases. Tumor microenvironment components as immune cells contribute to the tumor progression; thereby, the modulation of immune cells to a pro-inflammatory and antitumoral profile could potentialize the immunotherapy, a suggested approach for high-risk patients. Preview studies showed the antitumoral potential of BJcuL, a C- type lectin isolated from Bothrops jararacussu venom. It was able to induce immunomodulatory responses, promoting the rolling and adhesion of leukocytes and the activation of neutrophils. Methods: SK-N-SH cells were incubated with conditioned media (CM) obtained during the treatment of neutrophils with BJcuL and fMLP, a bacteria-derived peptide highly effective for activating neutrophil functions. Then we evaluated the effect of the same stimulation on the co-cultivation of neutrophils and SK-N-SH cells. Tumor cells were tested for viability, migration, and invasion potential. Results: In the viability assay, only neutrophils treated with BJcuL (24 h) and cultivated with SK-N-SH were cytotoxic. Migration of tumor cells decreased when incubated directly (p 0.001) or indirectly (p 0.005) with untreated neutrophils. When invasion potential was evaluated, neutrophils incubated with BJcuL reduced the total number of colonies of SK-N-SH cells following co-cultivation for 24 h (p 0.005). Treatment with CM resulted in decreased anchorage-free survival following 24 h of treatment (p 0.001). Conclusion: Data demonstrated that SK-N-SH cells maintain their migratory potential in the face of neutrophil modulation by BJcuL, but their invasive capacity was significantly reduced.

Neutrophils activated by BJcuL, a C-type lectin isolated from Bothrops jararacussu venom, decrease the invasion potential of neuroblastoma SK-N-SH cells in vitro

Neuroblastoma is a pediatric tumor with a mortality rate of 40% in the most aggressive cases. Tumor microenvironment components as immune cells contribute to the tumor progression; thereby, the modulation of immune cells to a pro-inflammatory and antitumoral profile could potentialize the immunotherapy, a suggested approach for high-risk patients. Preview studies showed the antitumoral potential of BJcuL, a C- type lectin isolated from Bothrops jararacussu venom. It was able to induce immunomodulatory responses, promoting the rolling and adhesion of leukocytes and the activation of neutrophils. Methods: SK-N-SH cells were incubated with conditioned media (CM) obtained during the treatment of neutrophils with BJcuL and fMLP, a bacteria-derived peptide highly effective for activating neutrophil functions. Then we evaluated the effect of the same stimulation on the co-cultivation of neutrophils and SK-N-SH cells. Tumor cells were tested for viability, migration, and invasion potential. Results: In the viability assay, only neutrophils treated with BJcuL (24 h) and cultivated with SK-N-SH were cytotoxic. Migration of tumor cells decreased when incubated directly (p < 0.001) or indirectly (p < 0.005) with untreated neutrophils. When invasion potential was evaluated, neutrophils incubated with BJcuL reduced the total number of colonies of SK-N-SH cells following co-cultivation for 24 h (p < 0.005). Treatment with CM resulted in decreased anchorage-free survival following 24 h of treatment (p < 0.001). Conclusion: Data demonstrated that SK-N-SH cells maintain their migratory potential in the face of neutrophil modulation by BJcuL, but their invasive capacity was significantly reduced.(AU)

Impact of topical nifedipine on wound healing in animal model (pig) / Impacto da nifedipina tópica na cicatrização de feridas em modelo animal (porco)

Abstract Background The human skin is an extremely sophisticated and evolved organ that covers the whole body. External agents or the patient's own diseases can cause skin injuries that can challenge healthcare professionals and impose high social, economic and emotional costs. Objectives To evaluate the impact of topical nifedipine on skin wound healing, specifically on polymorphonuclear cells, vascular proliferation, and collagen. Methods We used three pigs, and created eight injuries in the dorsal region of each animal. We applied 1%, 10%, and 20% concentration nifedipine creams to four of the wounds in animals 1, 2, and 3 respectively and treated the other twelve wounds with saline solution 0.9% only. We analyzed the presence of polymorphonuclear cells, vascular proliferation, and collagen at six different times (days 1, 3, 7, 14, 21, and 28). Results The evaluation of polymorphonuclear levels showed mild cell activity at all times in the control group, while in the nifedipine groups, marked levels were more frequent at all times during the experiment. There was a 4.84-fold increase in the chance of marked vascular proliferation (p = 0.019) and, at the same time, a decrease in collagen formation (OR 0.02 / p = 0.005) in animal 3. Conclusions Topical NFD may have an impact on skin wound healing mechanisms. Our study showed that polymorphonuclear cells and vascular proliferation increased. We also demonstrated that collagen formation decreased. Therefore, topical NFD may have a positive impact on skin wound healing. Additional studies are needed to confirm our results.

Resumo Contexto A pele humana é um órgão extremamente sofisticado e evoluído que cobre todo o corpo. As lesões cutâneas podem ser causadas por agentes externos ou pelas próprias doenças do paciente, e podem representar um desafio para os profissionais de saúde com altos custos sociais, econômicos e emocionais. Objetivos Avaliar o impacto da nifedipina tópica na cicatrização de feridas cutâneas, especialmente em relação a polimorfonucleares, proliferação vascular e colágeno. Métodos Utilizamos três porcos e realizamos oito ferimentos na região dorsal de cada animal. Aplicamos as concentrações de nifedipina creme a 1%, 10% e 20% para os animais 1, 2 e 3, respectivamente, sendo que, em quatro ferimentos, aplicamos o creme e, nos outros quatro ferimentos, apenas soro fisiológico a 0,9%. Analisamos a presença de polimorfonucleares, proliferação vascular e colágeno em seis momentos diferentes (dias 1, 3, 7, 14, 21 e 28). Resultados A avaliação dos níveis polimorfonucleares mostrou atividade celular discreta em todos os momentos no grupo controle, enquanto nos grupos nifedipina, os níveis marcados foram mais frequentes em todos os momentos do experimento. Houve aumento de 4,84 vezes na chance de uma produção marcada (p = 0,019) da proliferação vascular e, ao mesmo tempo, diminuição da formação do colágeno (odds ratio, OR 0,02; p = 0,005) no animal 3. Conclusões A nifedipina tópica pode ter impacto no mecanismo de cicatrização cutânea. Nosso estudo mostrou que há aumento dos polimorfonucleares e da proliferação vascular. Além disso, há diminuição da formação do colágeno. Assim, a nifedipina tópica pode ter impacto positivo na cicatrização das feridas cutâneas. Estudos adicionais são necessários para confirmar nossos resultados.

Effect of PMNs and MMP-9 on corneal injury after alkali burn / 国际眼科杂志(Guoji Yanke Zazhi)

@#AMI: To study the relationship between the infiltration of polymorphonuclear neutrophils(PMNs)and the expression of matrix metalloproteinases 9(MMP-9)in corneal stroma injury after alkali burn.<p>METHODS: Cornea alkali-burned model was made in 25 rabbits, then animals were grouped and sacrificed at 3d, 7d, 14d, 21d and 28d, respectively. The condition developing of alkali-burned cornea was observed by slit lamp biommicroscopy. The corneas were enucleated for histopathological examination. The infiltration of PMNs was identified by hematoxylin eosin(HE)staining and the expression of MMP-9 was identified by immunohistochemisty in different periods.<p>RESULTS: The quantity of PMNs and MMP-9 increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. The area and depth of corneal stroma after alkali burn were the most severe on the 14d.<p>CONCLUSION: During the wound healing process, alkali-burned cornea has close relation with the infiltration of PMNs and the expression of MMP-9. The infiltration of PMNs and the expression of MMP-9 is positively correlated in corneal stroma injury after alkali burn.

Samsum ant venom modulates the immune response and redox status at the acute toxic dose in vivo

Background:Ant venoms express surface molecules that participate in antigen presentation involving pro- and anti-inflammatory cytokines. This work aims to investigate the expression of MHC-II, CD80 and CD86 on the polymorphonuclear cells (PMNs) in rats injected with samsum ant venom (SAV).Methods:Rats were divided into three groups - control, SAV-treated (intraperitoneal route, 600 μg/kg), and SAV-treated (subcutaneous route, 600 μg/kg). After five doses, animals were euthanized and samples collected for analysis.Results:The subcutaneous SAV-trated rats presented decreased levels of glutathione with increased cholesterol and triglyceride levels. Intraperitoneal SAV-treated animals displayed significantly reduced concentrations of both IFN-γ and IL-17 in comparison with the control group. However, intraperitoneal and subcutaneous SAV-treated rats were able to upregulate the expressions of MHC-II, CD80 and CD86 on PMNs in comparison with the control respectively. The histological examination showed severe lymphocyte depletion in the splenic white pulp of the intraperitoneal SAV-injected rats.Conclusion:Stimulation of PMNs by SAV leads to upregulation of MHC-II, CD 80, and CD 86, which plays critical roles in antigen presentation and consequently proliferation of T-cells. Subcutaneous route was more efficient than intraperitoneal by elevating MHC-II, CD80 and CD86 expression, disturbing oxidative stability and increasing lipogram concentration.

Cytological endometritis in dairy cows: diagnostic threshold, risk factors, and impact on reproductive performance

We determined the threshold proportion of polymorphonuclear leukocytes (PMNs) for a diagnosis of cytological endometritis (CEM), the risk factors for this condition, and its impact on reproductive performance in dairy cows. Uterine cytology was performed on 407 Holstein cows 4 weeks postpartum to determine the proportions of endometrial cells and PMNs. A receiver operator characteristics curve was used to determine the threshold above which the PMN proportion affected the likelihood of cows conceiving by 200 days postpartum. The optimal threshold was ≥ 14% PMN (sensitivity, 31.3%; specificity, 81.7%; p < 0.05). The farm identity, retained placenta (odds ratio [OR] = 1.87), and septicemic metritis (OR = 3.07) were risk factors for CEM (p < 0.05). Cows with CEM were less likely to resume cyclicity (OR = 0.58) and to conceive by 200 days postpartum (hazard ratio = 0.58). Cows with CEM tended (p < 0.1) to be less likely to become pregnant after their first insemination (OR = 0.65) and to require a greater number of inseminations per conception (2.3 vs. 2.2). In conclusion, a PMN threshold of 14% defined the presence of CEM at 4 weeks postpartum. The farm, retained placenta, and septicemic metritis were risk factors for CEM, which reduces subsequent reproductive performance.

Comparison of CSF analysis by manual counts and automated hematology analyzer Sysmex XN-2000 / Эрүүл мэндийн лаборатори

Introduction:When a central nervous system disorder (meningitis, encephalitis, hemorrhage, leukemia infltration and other neoplasma) is present, cerebrospinal fluid (CSF) shows various changes that reflected the condition. Therefore it is essential to test CSF. Different types of CSF tests include cell count; cell differentiation; chemistry; immunology; microbiology and molecular biology. CSF cell count and cell differentiation in particular, are crucial in differentiating diagnosing various CNS disorder needing immediate care and in evaluating the treatment. The patient’s prognosis largely depends on how accurate diagnosis was done and how early treatment was provided. There for CSF test require high precision and accuracy. In Mongolia until now 2st and 3st level hospital using manual method for CSF cell count and cell differentiation test. In this test has 2 actual problems, which is depends on the analytical techniques, skills and sample stability specific problem. But in Japan in 2011 newly designed Sysmex XN Series hematology analyser with body fluid mode (CSF,pleural effusion, peritoneal and synovial fluid). On The First Central Hospital of Mongolia In 2013 frst timeinstalled Sysmex XN-2000 hematology analyser andpossible use of body fluid automatic testing methods.Materials and methods:We evaluated the basic assay performance of the body fluid mode on the automated hematology analyzer XN-2000, which is used for analysis of CSF fluid. We compared between the manual method and XN-2000 analysis for nucleated (WBC), mononuclear (MN) and polymorphonuclear (PMN) cells was also randomly studied using 10 CSF samples of inpatient section our hospital.Results:In CSF samples the coeffcient correlation(r) for WBC/µl, MN%, PMN% were respectively 0.83, 0.95 ба 0.95.Discussion:The correlation for MN%, PMN% were between automate and manual method was good, that is similar to the other researchers. Whereas the correlation for WBC/µl slightly low, this was probably correlation relatively weak or show discrepancies. In introduction inscriptive in analysis accuracy can to affect analytical techniques skills, sample stability and specifc many problems. Therefore scientifc studied and proven ability specifcity, sensitivity, reproducibility, quality, personnel low cost and spend less time, automatically Sysmex XN series hematology analyzer is desirable to domesticate an appropriate level of medical laboratories.

Changes in infiltration of polymorphonuclear neutrophils in alkali-burned cornea of conjunctival flap covering / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To study the infiltration of polymorphonuclear neutrophils ( PMNs ) after conjunctival flap covering in alkali-burned cornea. ●METHODS: Rabbit cornea alkali-burned model was made, then 50 rabbits were randomly divided into the experimental group ( n=25 ) and the control group ( n=25 ) . At the same time the surgery of conjunctival flap covering was given to rabbits of the experimental group. The condition developing of alkali-burned cornea was observed by slit lamp biomicroscopy, and took photos in two groups. The infiltration of PMNs was identified by hematoxylin eosin ( HE) staining in different periods. ●RESULTS:The quantity of PMNs increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. PMNs level of the experimental group was significantly lower than that in the control group, and the difference of 3, 14 and 21d was significant (P ●CONCLUSION: During the wound healing process, alkali - burned cornea has close relation with the infiltration of PMNs. The treatment of conjunctival flap covering for the severe alkali-burned cornea was found to have good effect.

Expression of CD14 and toll-like receptors 2 and 4 by milk neutrophils in bovine mammary glands infected with Corynebacterium bovis / Expressão de CD14 e dos receptores do tipo toll 2 e 4 por neutrófilos lácteos provenientes de glândulas infectadas por Corynebacterium bovis

This study evaluated the expression of CD14, toll-like receptor (TLR) 2 and TLR4 on the surface of milk neutrophils in bovine mammary glands infected with Corynebacterium bovis. Here, we used 23 culture-negative control quarters with no abnormal secretion on the strip cup test and milk somatic cell count lower than 1x105 cells/mL, and 14 C. bovis infected quarters. The identification of neutrophils, as well as, the percentage of neutrophils that expressed CD14, TLR2 and TLR4 were analyzed by flow cytometry using monoclonal antibodies. The present study encountered no significant difference in the percentages of milk neutrophils that expressed TLR2 and TLR4 or in the expression of TLR4 by milk neutrophils. Conversely, a lower median fluorescence intensity of TLR2 in milk neutrophils was observed in C. bovis-infected quarters. The percentage of neutrophils that expressed CD14 and the median fluorescence intensity of CD14 in milk neutrophils was also lower in C. bovis-infected quarters...

O presente estudo objetivou avaliar alterações na expressão de CD14, e dos receptores do tipo toll (TLR) 2 e 4 na superfície de neutrófilos lácteos provenientes de glândulas mamárias infectadas por Corynebacterium bovis. O presente estudo utilizou 23 quartos negativos no exame bacteriológico, sem alterações na prova de fundo escuro e com contagem automática de células somáticas menor que 1 x105 células/mL, e 14 quartos mamários infectados por C. bovis A identificação de neutrófilos, assim como a porcentagem de neutrófilos lácteos que expressaram CD14, TLR2 e 4 foram avaliadas por citometria de fluxo utilizando anticorpos monoclonais. A porcentagem de neutrófilos que expressaram TLR2 e TLR4 nos quartos mamários infectados por C. bovis não diferiu dos quartos mamários sadios, assim como na expressão de TLR4. No entanto, a intensidade de fluorescência do TLR2 na superfície dos neutrófilos foi menor nos quartos mamários infectados por C. bovis. A porcentagem de neutrófilos que expressaram CD14 e a intensidade de fluorescência da molécula de CD14 foi menor na superfície dos neutrófilos lácteos dos quartos infectados por C. bovis...

The effect of concentration ratio on tumor cell adhesion in shear flow / 医用生物力学

Objective To study the different effects from different concentration ratios of polymorphonuclear neutrophil (PMN) to tumor cell (TC) on the process of tumor cell adhesion to endothelial cell (EC) in shear flow. Methods PMNs and TCs with different concentration ratios (PMN-TC ratio) were added into the parallel plate flow chamber, and changes in the numbers of transient and accumulative adhered TCs on ECs at different shear rates (50 s-1，100 s-1，200 s-1) were analyzed. Results The transient and accumulative adhesion of TCs on ECs at PMN-TC ratio of 3︰1 significantly increased as compared to that at PMN-TC ratio of 1︰1, especially under high shear flow condition (100 s-1 and 200 s-1). Moreover, in the 5 minute-observation period, the effect of PMN-TC ratio on TC adhered to ECs occurred earlier when the shear rate increased. Conclusions The increase of PMN-TC concentration ratio can promote TC adhesion to ECs in shear flow, and the research findings provide significant references for studying TC metastasis in blood vessels and the target therapy of tumors.

Clinical value of neutrophil gelatinase -associated lipocalin and lactate dehydrogenase detection for screening of bacterial peritonitis in patients with ascites / 中国基层医药

Objective To build a simple,automatable detection method for diagnosing bacterial peritonitis. Methods Neutrophil gelatinase -associated lipocalin(NGAL),lactate dehydrogenase(LDH),proteins and glucose were assessed in peritoneal fluids from patients with nonmalignant ascites,along with nucleated cell count and differ-entiation.Results One hundred and eleven specimens were analyzed,26 of which with PMN count≥250 ×106 /L, thus compatible with bacterial peritonitis.The median concentration of LDH and NGAL was 3.4 and 3.7 -fold higher in samples with PMN≥250 ×106 /L.The concentration of proteins was also higher in samples with ≥250 ×106 PMN /L, whereas that of glucose was lower.The PMN count significantly correlated with peritoneal fluid values of LDH(r =0.859),NGAL(r =0.774)and proteins(r =0.268),but not with glucose(r =-0.069).The area under ROC curve was 0.88 for LDH,0.89 for NGAL and 0.94 for their combination(both tests positive),whereas that of proteins and glucose was 0.80 and 0.71,respectively.Sensitivities and specificities were 0.81 and 0.87 for LDH≥227U /L, 0.96 and 0.75 for NGAL≥120μg/L,0.77 and 0.95 for their combination.The agreement with PMN count was 0.86 for LDH,0.80 for NGAL,and 0.91 for their combination.Conclusion Assessment of NGAL in peritoneal flu-ids,especially in combination with LDH,may be a reliable approach for screening of bacterial peritonitis in patients with nonmalignant ascites.

Effect of dexmedetomidine on apoptosis and respiratory burst of polymorphonuclear neutrophils in vitro / 实用医学杂志

Objective To investigate the effect of dexmedetomidine (Dex) on the apoptosis and respiratory burst of human polymorphonuclear neutrophils (PMN) in vitro. Methods The peripheral venous blood was collected from healthy volunteers. Isolation of PMN was performed by using the discontinuous plasma-Percoll gradient technique. PMN was randomLy divided into four groups: control group and three different concentrations of Dex (1 ng/mL,10 ng/mL,100 ng/mL) groups. PMN was cultured in enriched RPMI-1640 media at 2 × 106/mL for 24 h. The caspase-3 activity, apoptosis rate and respiratory burst of PMN were detected at 2 and 24 h. Results Compared with the control group, the PMN which was treated with three different concentrations of Dex showed no significant difference in caspase-3 activity, apoptosis rate and respiratory burst of PMN after 2 h of incubation. Compared with the control group, 1 ng/mL concentration of Dex did not affect the caspase-3 activity, apoptosis and respiratory burst of PMN cultured for 24 h. However, 10 ng/mL and 100 ng/mL concentrations of Dex promoted the caspase-3 activity, increased apoptosis rate and reduced the respiratory burst of PMN after 24 h of incubation. Compared with 10 ng/mL Dex group, 100 ng/mL concentrations of Dex promoted the caspase-3 activity, increased apoptosis rate and reduced the respiratory burst of PMN after 24 h of incutation. Conclusion Clinically relevant concentration of Dex does not affect apoptosis and respiratory burst of PMN, while high concentration of dexmedetomidine can induce apoptosis of PMN.

Characteristics of cerebrospinal fluid in neonatal purulent meningitis / 中国小儿急救医学

Objective To analyze the changes of WBC classification,sugar and protein in cerebro-spinal fluid(CSF)and pathogenic bacteria of neonatal purulent meningitis.Methods Thirty-one neonates with bacterial meningitis in our department of neonatology from June 201 1 to June 2013 were enrolled,and the clinical features,pathogenic bacteria,laboratory examination of CSF were analyzed.Results Fever (90.3%),convulsions(67.7%)and changed consciousness(58.1 %)were common clinical symptoms.The incidences of other nervous system abnormal signs such as gastrointestinal dysfunction(25.8%),abnormal breathing(16.1 %),cervical resistance(16.1 %),bulging fontanel(9.7%)were lower.The Gram -negative bacteria was more than Gram -positive in both blood and CSF culture.The escherichia coli was the most common bacteria,with positive rate of 38.1 %(8 /21 )in blood culture and 55.5%(5 /9)in CSF culture.The germiculture positive rate in CSF was lower than in blood culture (29.0% vs.67.7%).Polymorphonuclear leukocyte(PMN)[(79.61 ±12.06)%]was the most predominant cell of the leukocyte classification in CSF within 1 week in all cases,PMN was still predominant in 1 to 2 weeks in 7 cases,while only 2 cases in 2 to 3 weeks still dominated by PMN,PMN was not the predominant cell 3 weeks later.Conclusion In the typi-cal neonatal purulent meningitis,PMN was the predominant cell in CSF within the first week,but the propor-tion of monocyte gradually increased and was dominant later.Escherichia coli was a common bacteria caused by this disease.

Células inflamatorias en la secreción nasal y citocinas proinflamatorias Th1, Th2, Th17 y reguladoras en el suero de pacientes con leishmaniasis cutánea Americana / Inflammatory cells in the nasal secretion and serum pro-inflammatory Th1, Th2, Th17 and regulatory cytokines from American cutaneous leishmaniasis patients

En la forma mucocutánea (LCM) y cutánea (LCL) de la leishmaniasis, se genera una respuesta inflamatoria cuyos mediadores (células y citocinas) se han involucrado en la severidad de las úlceras y en el daño tisular observado en estos pacientes, particularmente en los LCM. Por ello, nos propusimos identificar los grupos celulares predominantes en la secreción nasal de pacientes con LCL y LCM, y relacionarlos con citocinas proinflamatorias y reguladoras. Evaluamos en pacientes LCL (n=20), LCM (n=14) y 20 individuos sanos: a) La cuantificación de tipos de leucocitos en "frotis" de secreción nasal, úlceras cutáneas y sangre periférica teñidos con Giemsa empleando microscopía óptica, b) Concentraciones séricas de IL-8, IL-4 e IL-10 por citometría de flujo (CBA array) e IFN-γ, TNF-α e IL-17 por ELISA. El grupo celular predominante en la secreción nasal de pacientes con LCM fueron los neutrófilos (80,7%) y escasos eosinófilos (0,6%), comparados con los LCL y controles, en los que no se observaron estas células. Mientras que los "frotis" de las ulceras de los LCL presentaron 45,3% de neutrófilos y 43% de linfocitos. En contraste, en sangre periférica, de los pacientes se observó un incremento de neutrófilos y linfocitos junto a una frecuencia significativa de monocitos (LCM: 5,3; LCL: 6,3%) y eosinófilos (LCM: 8,2%; LCL: 5,2%). Todo esto sugiere la participación de los neutrófilos en la inmunopatogénesis en la LCM. Adicionalmente, se demostró una mayor (P=0,03) concentración sérica de IL-8 en los pacientes con LCL (18,5ρg/mL) y LCM (18,2ρg/mL) respecto a los individuos sanos, sugiriendo que esta citocina promueve el reclutamiento de neutrófilos al sitio de infección en los LCM, mientras que en los LCL contribuyen junto con los linfocitos T CD4+ de la subpoblación Th1 y productores de IFN-γ, en la activación de mecanismos leishmanicidas.

In mucocutaneous (MCL) and cutaneous (LCL) leishmaniasis, the inflammatory mediators (cytokines and cells) have been associated with ulcers severity and tissue damage observed in these patients, particularly in MCL. Therefore, we decided to identify the predominant cell groups in the nasal secretion of LCL and MCL patients, and related pro-inflammatory and regulatory cytokines. It was evaluated in LCL (n = 20), MCL patients (n = 14) and 20 healthy volunteers: a) Differential leukocyte count by optical microscopy performed in: smear of a runny nose, skin ulcers and peripheral blood dyed with Giemsa, b) serum levels of IL-8, IL-4 and IL-10 using cytometric bead array (CBA) assay and IFN-γ, TNF-α and IL-17 by ELISA. In MCL patients, neutrophils (80.7%) were the most abundant cellular group in nose secretion, followed by a small amount of eosinophils (0.6%) compared to the LCL and controls, where no such cells were observed. In contrast, in peripheral blood from ACL patients were observed an abundant amount of neutrophils and lymphocytes together with a significant frequency of monocytes (MCL:5.3%; LCL: 6.3%) and eosinophils (MCL:8.2%; LCL:5.2%). While the smear from skin ulcers of LCL patients showed 45.3% of neutrophils and 43% lymphocytes. All of these indicate that neutrophils might play a role in the MCL immunopathogenesis. Moreover, an increased serum levels of IL-8 (P=0.03) were found in LCL (18.5ρg/mL) and MCL (18.2ρg/mL) patients, suggesting that this cytokine promotes the recruitment of neutrophils to the infection site in MCL; while in LCL patients may contribute with CD4 + Th1 (IFN-γ) cells in the activation of leishmanicida mechanisms.

Effects of Fruits of Barringtonia Racemosa Linn. On Human Polymorphonuclear Cell.

The objective of present study was to investigate Petroleum ether, ethyl acetate and hydroalcoholic extracts of B. racemosa fruits in vitro on human polymorphonuclear (PMN) cells to screen their effects on phagocytosis and chemotaxis. Ethyl acetate extract of B. racemosa fruitswas found to be a stimulant of PMN cell phagocytosis of Nitroblue tetrazolium (NBT) dye and candida albicans. It also stimulated intracellur killing capacity of PMN cells. It was further found to increase the chemotaxis of human PMN cells.While, petroleum ether extract and hydroalcoholic extract were lesser active as far as these activities are concerned.

Efectos de extractos de Neopetrosia rosariensis sobre mieloperoxidasa y elastasa con la utilización del modelo neutrófilos / Effects of extracts from Neopetrosia rosariensis on myeloperoxidase and on elastase by using the neutrophil model

Introducción: los neutrófilos son la primera línea de defensa, los cuales liberan su contenido granular, constituido entre otros por enzimas generadoras de especies reactivas del oxígeno como mieloperoxidasa y enzimas proteolíticas como la elastasa, que degradan elastina y colágeno. Objetivo: evaluar la toxicidad de los extractos de la esponja marina Neopetrosia rosariensis y su acción como inhibidores de la desgranulación del neutrófilo, e inhibición de la actividad catalítica de la elastasa de neutrófilos humanos. Métodos: se determinó la toxicidad por el método de exclusión del colorante azul de tripano. Se evaluó la desgranulación por la disminución del porcentaje de liberación de la mieloperoxidasa, y se determinó la inhibición de la actividad catalítica de la elastasa de neutrófilos humanos por la reducción de la actividad de la enzima de un sustrato específico. Resultados: los extractos presentaron baja toxicidad a las concentraciones evaluadas, con porcentaje de viabilidad celular superior al 80 por ciento, incluso a la concentración más alta utilizada (25 µg/mL). También presentaron actividad inhibitoria de la liberación de mieloperoxidasa, concentrándose el efecto en los extractos más polares (metanólico total y parcial) y la inhibición de la actividad de la elastasa en los de más baja polaridad (hexano y diclorometano parcial), a concentraciones de 10 y 25 µg/mL, acorde con la naturaleza lipofílica del sitio activo de la enzima. Conclusiones: los resultados en los extractos evaluados son promisorios, teniendo en cuenta que cada muestra representa una mezcla compleja de compuestos activos entre 10 y 25 µg/mL, lo que resulta de mucho interés para continuar con su fraccionamiento como agentes potenciales para la posible terapia de enfermedades que presentan procesos inflamatorios(AU)

Introduction: neutrophils are the first line of defense, releasing their granular contents made up of reactive oxygen species-generating enzymes such as myeloperoxidase and proteolytic enzymes such as elastase, which degrade elastin and collagen. Objective: to evaluate toxicity of marine sponge Neopetrosia rosariensis extracts and their action as inhibitors of neutrophil degranulation and inhibition of the catalytic activity of human neutrophil elastase. Method: the cytotoxicity was assessed by Trypan blue exclusion test of cell viability. Degranulation inhibition was evaluated by reduction of myeloperoxidase release percentage and inhibition of catalytic activity of elastase was determined by reduction of enzyme activity on specific substrate. Results: the extracts exhibited low toxicity at the evaluated concentrations, with cellular viability percentage exceeding 80 percent even at the highest concentration (25 µg/mL). They also showed inhibitory action of myeloperoxidase release, the greatest effect being concentrated on the most polar extracts (total and partial methanolic). The inhibition of the elastase activity was mostly found in the lowest polarity extracts (hexane and partial dichloromethane) at concentrations of 10 and 25 µg/mL, according to the hydrophobic character of the active site of the enzyme. Conclusions: these results of the evaluated extracts are encouraging, considering each extract represents a complex mixture of active compounds of 10-25 µg/mL. This arouses great interest to continue their fractioning as potential agents for possible therapy of inflammatory processes(AU)

Correlation between Percentages of PMN, MNC, FBC and Wound Age after Skeletal Muscle Injury in Rats / 法医学杂志

Objective To study the percentages of polymorphonuclear leukocytes (PMN), mononuclear cells (MNC) and fibroblastic cells (FBC) in different post-traumatic intervals after skeletal muscle me-chanical injury in rats. Methods The rat model of skeletal muscle mechanical injury was established. The rats were divided into injured groups (6 h, 12 h, 1 d, 3 d, 7 d, 10 d and 14 d after injury ) and con-trol group. The percentages of PMN, MNC and FBC in different post-traumatic intervals after skeletal muscle mechanical injury were assessed with HE staining and image analysis. Results At post-injury 6-12 h, the percentages of PMN and MNC infiltration appeared in injured sites and that of PMN reached peak. At 1 d, the percentage of MNC infiltration appeared and reached peak, while that of PMN de-creased. At 3-7 d, the percentage of FBC gradually increased, while that of PMN and MNC decreased. At 10-14 d, the percentage of FBC reached peak. Conclusion The percentages of PMN, MNC and FBC in injured zones showed time-dependent changes, which might be used as reference index for determination of age of skeletal muscle injury.