Efficacy of minocycline hydrochloride ointment combined with metronidazole film in the treatment of periodontitis and their effects on C-reactive protein and elastase levels in gingival crevicular fluid / 中国基层医药

Objective:To investigate the efficacy of minocycline hydrochloride ointment combined with metronidazole film in the treatment of periodontitis and their effects on C-reactive protein and elastase levels in the gingival crevicular fluid.Methods:76 patients with periodontitis who received treatment in Jiaxing Hospital of Traditional Chinese Medicine from May 2019 to January 2020 were included in this study. They were randomly allocated to undergo treatment with metronidazole film (control group, n = 38) or minocycline hydrochloride ointment plus metronidazole film (observation group, n = 38) for 4 weeks. We compared clinical efficacy, periodontal system examination indexes (gingival index, periodontal probing pocket depth, gingival bleeding index, plaque index, loss of attachment), gingival crevicular fluid biochemical markers (C-reactive protein, elastase in the pellet, elastase in the supernatant) measured before and after treatment, the incidence of adverse reactions, and the recurrence rate within half a year after treatment between the two groups. Results:The total response rate was significantly higher in the observation group than in the control group [97.37% (37/38) vs. 78.95% (30/38), χ2 = 6.17 , P < 0.05]. Gingival index, periodontal probing pocket depth, gingival bleeding index, plaque index, and loss of attachment measured after treatment were significantly lower in the observation group than in the control group (all P < 0.001). C-reactive protein, elastase in the pellet, and elastase in the supernatant measured after treatment were (5.31 ± 1.19) μg/L, (0.70 ± 0.20) Abs/mL, (0.48 ± 0.19) Abs/mL respectively in the observation group, which were significantly lower than those in the control group [(7.92 ± 1.27) μg/L, (1.15 ± 0.52) Abs/mL, (1.12 ± 0.31) Abs/mL, t = 9.24, 4.97, 10.85, all P < 0.001]. The recurrence rate within half a year in the observation group was significantly lower than that in the control group [2.63% (1/38) vs. 20% (6/38), χ2 = 3.93, P < 0.05]. There was no significant difference in the incidence of adverse reactions between the two groups ( P > 0.05). Conclusion:Minocycline hydrochloride ointment combined with metronidazole film is safe and effective in the treatment of periodontitis. The combined therapy help downregulate the levels of C-reactive protein, elastase in the pellet, elastase in the supernatant of the gingival crevicular fluid, alleviate inflammation, improve the periodontal status, and reduce the recurrence rate.

Elastase-Positive Neutrophils Are Associated With Refractoriness of Chronic Rhinosinusitis With Nasal Polyps in an Asian Population

PURPOSE: Various immune cells, including eosinophils and neutrophils, are known to contribute to the development of chronic rhinosinusitis with nasal polyps (CRSwNP). However, the current understanding of the role of neutrophils in the development of CRSwNP still remains unclear. Therefore, we investigated risk factors for refractoriness of CRSwNP in an Asian population. METHODS: Protein levels of 17 neutrophil-related mediators in nasal polyps (NPs) were determined by multiplex immunoassay, and exploratory factor analysis using principal component analysis was performed. Immunofluorescence analysis was conducted to detect human neutrophil elastase (HNE) or myeloperoxidase (MPO)-positive cells. Tissue eosinophilic nasal polyp (ENP) and tissue neutrophilia (Neu(high)) were defined as greater than 70 eosinophils and 20 HNE-positive cells, otherwise was classified into non-eosinophilic nasal polyp (NENP) and absence of tissue neutrophilia (Neu(low)). RESULTS: In terms of disease control status, NENP-Neu(low) patients showed the higher rate of disease control than NENP-Neu(high) and ENP-Neu(high) patients. Linear by linear association demonstrated the trend in refractoriness from NENP-Neu(low) to NENP-Neu(high) or ENP-Neu(low) to ENP-Neu(high). When multiple logistic regression was performed, tissue neutrophilia (hazard ratio, 4.38; 95% confidence interval, 1.76-10.85) was found as the strongest risk factor for CRSwNP refractoriness. Additionally, exploratory factor analysis revealed that interleukin (IL)-18, interferon-γ, IL-1Ra, tumor necrosis factor-α, oncostatin M, and MPO were associated with good disease control status, whereas IL-36α and IL-1α were associated with refractory disease control status. In subgroup analysis, HNE-positive cells and IL-36α were significantly upregulated in the refractory group (P = 0.0132 and P = 0.0395, respectively), whereas MPO and IL-18 showed higher expression in the controlled group (P = 0.0002 and P = 0.0009, respectively). Moreover, immunofluorescence analysis revealed that IL-36R⁺HNE⁺-double positive cells were significantly increased in the refractory group compared to the control group. We also found that the ratio of HNE-positive cells to α1 anti-trypsin was increased in the refractory group. CONCLUSIONS: Tissue neutrophilia had an influence on treatment outcomes in the Asian CRSwNP patients. HNE-positive cells and IL-36α may be biomarkers for predicting refractoriness in Asians with CRSwNP. Additionally, imbalances in HNE and α1 anti-trypsin may be associated with pathophysiology of neutrophilic chronic rhinosinusitis.

Cloning of Guamerin gene in Whitmania pigra and its spatio-temporal expression analysis after ingestion / 中国中药杂志

The present study cloned wpGuamerin gene from a non-bloodsucking leech (Whitmania pigra), and the bioinformatics analysis of the sequence was performed. Additionally, the effects of feeding duration on the expression profile of the wp Guamerin gene were explored. The results showed that its sequence consisted of 295 nucleotides encoding a peptide of 83 amino acids(Genbank: KX768545), and its relative molecular weight is 9 145.95 Da. wp Guamerin does not encode proteins with a signal peptide, belonging to the hydrophilic protein. Its secondary structure is mainly composed of α-helix, extending chain, folding and random curl. Its similarity with other blood-sucking leeches ranges from 29% to 65%. The results revealed that wpGuamerin mRNA was detected higher expression in muscle than in salivary glands of Wh. pigra, and did not expressed in ingluvies and intestine. Its expression in muscle and salivary glands showed a single peak curve after feeding and the peak was observed in the 1st and 3rd after feeding, respectively. In summary, wp Guamerin in Wh. pigra is a small molecule polypeptide protein and is different from the Guamerin in blood-sucking leeches. wpGuamerin does not express in the digestive tract of Wh. pigra, and mainly express in muscle. Feeding behavior would stimulate the expression of wpGuamerin gene in muscle and salivary glands, but not in digestive tract.

Effects of sivelestat sodium on early inflammatory reaction in rats with smoke inhalation injury / 天津医药

Objective To investigate the effects of sivelestat sodium on early inflammatory reaction in rats with smoke inhalation injury. Methods Forty SPF male SD rats were randomly divided into 5 groups:normal control group (A), injury group (B), smoke inhalation treated with 10 mg/kg sivelestat sodium group (C), smoke inhalation treated with 20 mg/kg sivelestat sodium group (D) and smoke inhalation treated with 30 mg/kg sivelestat sodium group (E), 8 rats for each group. After smoke inhalation injury model was established, the treatment groups were intraperitoneally injected sivelestat sodium 10 mg/kg, 20 mg/kg and 30 mg/kg separately. B group was treated with the same volume of physiological saline. After 24 hours,ELISA was used for detecting serum contents of neutrophil elastase (NE), myeloperoxidase (MPO), tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in five groups. Meanwhile the water content of lung tissue was measured, and the pathological changes were observed by HE staining. The thickness of alveolar septum was measured and compared between groups. Results Compared with control group, the serum levels of NE, MPO, IL-6, TNF-α, water content of the lung tissue and thickness of alveolar septum were significantly higher in other four groups (P<0.05). Compared with injury group, the serum levels of NE, MPO, IL-6, TNF-α, water content of the lung tissue and thickness of alveolar septum were significantly lower in treatment groups (P<0.05). Compared with 20 mg/kg treatment group and 30 mg/kg treatment group, the serum levels of NE, MPO, IL-6, TNF-α, water content of the lung tissue and thickness of alveolar septum were significantly lower in 10 mg/kg treatment group (P<0.05). Conclusion The result shows that sivelestat sodium can reduce the early inflammatory reaction of rats with smoke inhalation injury and attenuates the lung edema. In this experiment, the treatment effect of 10 mg/kg sivelestat sodium is better than other treatment doses.

Neutrophil elastase ameliorates matrix metalloproteinase-9 to promote lipopolysaccharide-induced acute lung injury in mice 1

ABSTRACT PURPOSE: To investigate the regulatory roles of neutrophil elastase (NE) and matrix metalloproteinase-9 (MMP-9) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. METHODS: To construct LPS-induced ALI mouse models, wild-type C57BL/6 mice were administered 5.0 mg/kg of LPS through endotracheal, and/or 1.0 mg/kg of ONO-5046, and/or 20.0 mg/kg of chemically modified tetracycline-3 (CMT-3) by gavage. The levels of MMP-9, tissue inhibitor of metalloprotease-1, interleukin (IL)-6 were detected by real time RT-PCR at 6 h, 24 h and 48 h, and tumor necrosis factor (TNF), lung wet-dry weight ratio, white blood cell (WBC) count and polymorphonuclear (PMN) count in bronchoalveolar lavage fluid (BALF) were tested at 48 h after administration. The 5-day survival analysis of the ALI mice was also performed. RESULTS: Both ONO-5046 and CMT-3, regardless of being used individually or combined, significantly reduced the levels of MMP-9, IL-6, and TNF in lung tissue as well as in BALF, and the WBC and PMN count in BALF. Combined treatment with ONO-5046 and CMT-3 remarkably improved the survival rate of ALI mice. CONCLUSION: Neutrophil elastase synergizes with matrix metalloproteinase-9 to promote and regulate the release of inflammatory mediators and the infiltration of inflammatory cells, consequently affecting the survival of lipopolysaccharide-induced acute lung injury mice.

Preventive effects of ulinastatin on acute respiratory distress syndrome / 北京大学学报(医学版)

Objective:To explore the effect of ulinastatin on prevention of acute respiratory distress syn-drome (ARDS).Methods:A prospective multicentral cohort study was conducted.The patients from three intensive care units (ICUs)of grade A tertiary hospitals in Beijing and a ICU of grade A tertiary hospitals in Cangzhou from January 2012 to December 2014,included 77 ARDS at-risk patients with uli-nastatin treatment and 108 ARDS at-risk patients without ulinastatin treatment (control)were eligible. Both groups received normal treatment;additionally,the intervention group received 600 000 units of uli-nastatin via intravenous infusion for 5 days.The control group received the same amount of saline via in-travenous infusion for 5 days.Venous blood human neutrophil elastase (HNE)and peptidase inhibitor 3 (PI3)levels were measured on days 1,3,and 7,respectively.Other outcomes included acute physiolo-gy and chronic health evaluation scoring Ⅱ (APACHE Ⅱ),body temperature,respiratory rate,heart rate,mean arterial pressure,white blood cell counts,PaO2 /FiO2 ,ARDS incident,mechanical ventila-tion time,ICU treatment and hospitalization duration,28 days mortality.Results:The PI3 levels showed no statistical difference on day 1,but significant differences on day 3 and day 7 between the two groups (P <0.01).HNE /PI3 ratio showed no statistical difference on day 1,but significant differences on day 3 and day 7 (P <0.05).PaO2 /FiO2 was significantly higher in ulinastatin group on day 3 and day 7 (P <0.05).The incident rate for ulinastatin group was 15.58%,lower than that for the control group (33.33%),and the difference was statistically significant (P <0.05).The mechanical ventilation time and ICU treatment time in ulinastatin group was shorter than that in the control group,and the difference was statistically significant (P <0.05).There were no significant effects in other factors.Conclusion:Increased dose of ulinastatin can recover the balance of HNE and its antagonist,lower the HNE’s damage to lungs,and further reduce the ARDS incident rate.

Measurement of neutrophil elastase, citric acid and pH in prostatic fluid of type Ⅲ prostatitis / 中华全科医师杂志

Objective To measure the levels of neutrophil elastase (NE),citric acid and pH in prostatic fluid of patients with prostatitis type Ⅲ.Methods Fifty two patients with prostatitis type ⅢA,98 patients with prostatitis type Ⅲ B and 105 health subjects were enrolled in the study.The levels of NE,citric acid and pH in prostatic fluid were measured ; the expressed prostatic secretion (EPS) routine and bacterial culture were examined,and National Institutes of Health (NIH)-chronic prostatitis symptom index (NIH-CPSI) was evaluated.The data were analyzed by independent samples t-test.Results WBC count,NE and citric acid concentrations,pH in ⅢA group were (17.9 ±5.4)/HP,(898 ±704) μg/L and (14.5 ± 1.7) ng/L,7.60 ±0.43,respectively; those in ⅢB group were (3.9 ±2.2)/HP,(93 ±76) μg/L and (21.9 ±3.8) ng/L,6.71 ±0.25,respectively; those in control group were (3.6 ±2.2)/HP,(86 ±57) μg/L and (22.5 ± 3.9)ng/L,6.48 ± 0.51,respectively.There were significant differences in NE and citrate concentrations,pH value in EPS between Ⅲ A and Ⅲ B patients (t =8.22,16.64 and 13.88,all P ＜0.05),but no difference in NIH-CPSI (t =1.90,P 8 0.05).There were significant differences in WBC count,NE and citric acid concentrations,pH in EPS,NIH-CPSI score between Ⅲ A and control groups (t =18.92,8.47,26.53,18.37 and 32.47,all P ＜ 0.05).There were no differences in WBC count,NE and citric acid concentration and pH value in EPS between Ⅲ B group and control group (t =1.38,1.55,1.02 and 1.21,respectively,all P 80.05),but there was significant difference in NIH-CPSI score between two groups (t =49.46,P ＜ 0.05).In EPS,NE concentration and WBC count were positively correlated with NIH-CPSI (r =0.819 and 0.698,respectively,all P ＜0.01),and citric acid was negatively correlated (r =-0.625,P ＜ 0.01) ; citrate was negatively correlated with WBC count,CPSI and pH value (r =-0.728,-0.644 and-0.817,all P ＜ 0.01).Conclusions The results indicate that the measurements of NE,citric acid and pH in EPS are of significant clinical value in patients with prostatitis type ⅢA and Ⅲ B.

Relato de um caso de neutropenia congênita grave em um lactente jovem / A case report of severe congenital neutropenia in a young infant

OBJETIVO:Relatar um caso de neutropenia congênita grave e alertar os pediatras sobre tal diagnóstico em pacientes jovens, com infecções recorrentes. DESCRIÇÃO DO CASO: Lactente jovem com 45 dias de vida, com história de febre alta, letargia, recusa alimentar e hemogramas repetidos com leucopenia importante à custa de polimorfonucleares. A hipótese diagnóstica foi confirmada pelo aspirado de medula óssea, que mostrou hipoplasia de série granulocítica e completa ausência de neutrófilos maduros. Foi introduzida antibioticoterapia de largo espectro e estimulador da formação de colônias de granulócitos. O paciente evoluiu para óbito em decorrência de complicações infecciosas após 21 dias de internação. COMENTÁRIOS: Trata-se de um lactente jovem, portador de uma rara desordem congênita que leva à intensa neutropenia, deixando-o vulnerável a infecções graves e potencialmente fatais. À internação, o paciente apresentava sinais e sintomas sugestivos de sepse, sendo introduzido antibioticoterapia de amplo espectro, necessária por se tratar de lactente jovem, neutropênico e febril. A hipótese diagnóstica se baseou na história clínica e nos leucogramas alterados, sendo posteriormente confirmada pelo aspirado de medula óssea. Foi introduzido o estimulador da formação de colônias de granulócitos, que geralmente é efetivo, porém, nesse caso, não houve sucesso e o paciente evoluiu para óbito devido à grave infecção.

OBJECTIVE:To report a case of severe congenital neutropenia and alert pediatricians about its diagnosis in young patients with recurrent infectious diseases. CASE DESCRIPTION: Young infant with 45 days of life, with a history of high fever, lethargy, poor feeding and repeated blood counts showing significant leucopenia due to a significant decrease of polymorphonuclear cells. The diagnosis was confirmed by bone marrow aspirate showing hypoplasia of the granulocytic series and complete absence of mature neutrophils. Treatment was started with broad-spectrum antibiotic therapy and granulocyte colony-stimulating factor, but the patient died due to infectious complications 21 days after hospital admission. COMMENTS: This is a young infant with a rare congenital disorder that leads to severe neutropenia and, therefore, susceptible to potentially fatal infections. In the hospital the infant showed signs and symptoms of sepsis. The diagnosis was based on the clinical history and the presence of repeated altered white cell counts and it was confirmed by bone marrow aspirate. Granulocyte colony-stimulating factor is generally effective, but, in this case, the patient died with a severe infection.

Identification and bioactivities analysis of Halomonas sp. from seawater / 第二军医大学学报

Objective: To determine the 16S rDNA sequences of 36 strains of Halomonas sp. Isolated from the seawater of the East China Sea and to study their bioactivities. Methods: The 16S rDNA sequences of all isolated strains were amplified by PCR, and the obtained sequences were subjected to phylogenetic analyses using MEGA4.1 and multiple sequence alignment with Clustal X1. 8 software. MTT assay was applied to assess the cytotoxic activities of their fermentation broth; ABTS and DPPH antioxidant models were applied to test the antioxidation activities of their fermentation broth. The HLE inhibition activities of the fermentation broth were tested by a stable HLE in hibitor screening model. Results: Analysis of 16S rDNA sequence showed that the 36 strains had a high affinity (96%-99%) to the Halomonas sp. Fermentation broth of the 36 strains showed different degrees of cytotoxic activities, antioxidation activities, and HLE inhibition activities. Eleven strains inhibited the growth of HepG2 cells, with the inhibition rate ranging (4.40±1.2)%to(24.90±3.5) %; 20 strains inhibited the growth of HL-60 cells, with the inhibition rate ranging (1.70±1.1)% to (50.90±4.2) %; 7 strains showed ABTS radical scavenging capacity, with the scavenging rate ranging (4.49±2.1) % to (58.43±4.4) %; 3 strains showed DPPH radical scavenging capacity, with the scavenging rate ranging (5.68±3.7) % to (59.06±3.2) %, and 3 strains showed HLE inhibition activities, with the inhibition rate ranging (12.71±1.81)% to (71.19±5.62) %. Conclusion: All the 36 Halomonassp. Strains show different degrees of cytotoxic effects, antioxidation activities, and HLE inhibition capacities. Several strains with high activities possess the potential for medicinal application.

Impacto da terapia mecânica não cirúrgica na atividade de elastase e volume do fluido gengival em indivíduos com periodontites crônica e agressiva generalizadas

O objetivo deste estudo foi avaliar o impacto do tratamento periodontal não cirúrgico sobre a atividade de elastase e o volume de fluido gengival nos pacientes portadores de periodontites crônica e agressiva generalizadas. Foram avaliados 18 pacientes com periodontite crônica e 11 com periodontite agressiva. Foram utilizados os parâmetros clínicos de avaliação da profundidade de bolsa à sondagem, nível de inserção e sangramento à sondagem. As amostras foram colhidas em cinco sítios mais profundos e em cinco sítios rasos com gengivite de cada paciente, antes e 90 dias após o término do tratamento. A amostra analisada antes e após o tratamento não apresentou diferenças significativas entre os grupos, exceto para a profundidade de bolsa à sondagem nos sítios rasos com gengivite (p = 0,039) e para o sangramento à sondagem (p = 0,021) nos sítios profundos após o tratamento. Após o tratamento, a elastase apresentou uma redução significativa nos sítios profundos, para as periodontites crônica (p = 0,012) e agressiva (p = 0,02). Houve significativa redução do volume de fluído gengival nos pacientes com periodontite crônica e agressiva, nos sítios rasos (p = 0,03 e p = 0,03) e profundos (p < 0,001 e p = 0,003), respectivamente, após o tratamento. Concluindo, os grupos com periodontites crônica e agressiva generalizadas comportaram-se de maneira semelhante frente à terapia mecânica não cirúrgica. Houve uma redução significativa do volume de fluído gengival e da atividade neutrofílica nos sítios profundos, associada a reduções significativas dos indicadores clínicos após o tratamento.

Neutrophil elastase and leukemia / 国际肿瘤学杂志

Neutrophil elastase is an important cytotoxicity molecule secreted by neutrophil. It plays a significant role in the development of leukemia. Since this protease can cleave the fusion protein PML-RARα,it may play a role in acute promyelocytic leukemia pathogenesis by facilitating the leukemogenic potential of PML-RARα. Otherwise, this protease might contribute to the clonal dominance of chronic myelognous leukemia by reducing the availability of growth factors for normal hematopoiesis.

The Anti-Inflammatory Effects of Ulinastatin in Trauma Patients with Hemorrhagic Shock

We investigated the use of ulinastatin in association with the suppression of polymorphonuclear leukocyte elastase (PMNE), tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6), and its effects on the prognosis of patients with traumatic hemorrhagic shock. Nineteen patients who visited the emergency department for traumatic hemorrhagic shock were enrolled. Eleven patients were randomly selected to receive a total of 300,000 IU of ulinastatin. Measurements of serum PMNE, TNF-alpha and IL-6 were taken before ulinastatin treatment at 24 hr, two days, three days, and seven days after admission. We compared the Systemic Inflammatory Response Syndrome scores, Multiple Organ Dysfunction Syndrome scores and Acute Physiology, age, Chronic Health Evaluation III scores of the control and ulinastatin groups. There were no significant differences in baseline values, laboratory data, treatment or mortality between the two groups. The serum PMNE levels in the ulinastatin group were lower than in the control group on the second hospitalized day. Serum TNF-alpha and IL-6 levels in the ulinastatin group decreased 24 hr after admission but had no significance. It is suggested that ulinastatin treatment could decrease the serum PMNE levels in trauma patients with hemorrhagic shock at 48 hr after treatment.

The Effect of Human Neutrophil Elastase on the Human Nasal Mucus Secretion / 대한이비인후과학회지

BACKGROUD AND OBJECTIVES: Human neutrophil elastase (HNE), secreted from neutrophil, is known to cause mucus hypersecretion in case of airway inflammation. The purpose of this study is to find out about the effect of HNE on the secretion of nasal mucosa. SUBJECTS AND METHOD: For in vitro study, inferior turbinate mucosa samples were obtained from patients who had septal deviation and turbinate hypertrophy. Short-term organ culture was done with and without HNE (100 to 400 ng/ml). Concentration of mucin in the culture media was measured by enzyme linked lectin assay (ELLA). In in vivo study, lavage fluid was obtained from patients with chronic rhinosinusits (n=11) and from the patients with septal deviation as control (n=10) to measure the concentration of mucin and HNE by ELLA and ELISA, respectively. RESULTS: The concentrations of HNE and mucin were significantly higher in the lavage fluid obtained from patients with chronic rhinosinusits than in that obtained from control (HNE ; 26.6+/-14.6 : 9.7+/-5.5 microgram/ml, p=0.01. Mucin ; 56.5+/-9.3 : 29.1+/-15.5 ng/ml, p=0.0002). The correlation coefficient between the concentration of HNE and mucin was not statistically significant (r=0.423). The concentration of mucin increased in dose-dependent fashion by increasing the concentration of HNE in in vitro study and HNE (400 ng/ml) significantly increased the concentration of mucin compared with that of control. CONCLUSION: From this study, we suggest that increased HNE in the inflammation of nasal cavity could have important role in mucus hypersecrtion.

Neutrophil Elastase Causes MUC5AC Mucin Synthesis Via EGF Receptor, ERK and NF-kB Pathways in A549 Cells

BACKGROUND: Neutrophil elastase (NE) was found to increase the respiratory mucin gene, MUC5AC, although the molecular mechanisms of this process remain unknown. We attempted to determine the signal transduction pathway through which NE induces MUC5AC gene expression in bronchial epithelial cells. METHODS: A fragment of 1.3 Kb MUC5AC promoter which had been cloned into the pGL3-Basic luciferase vector was transfected to the A549 cells. By measuring the luciferase activity, we were able to evaluate the MUC5AC promoter activity in A549 cells. The involvement of mitogen-activated protein kinases (MAPK) was confirmed by Western blotting. To confirm the involvement of nuclear factor kappaB (NF-kB), we used site-directed mutagenesis and electrophoretic mobility shift assay (EMSA) autoradiogram. The MUC5AC mRNA expression was confirmed by RT-PCR. RESULTS: NE increased the transcriptional activity of the MUC5AC promoter in A549 cells. The increased transcriptional activity of the MUC5AC promoter by NE was found to be associated with increased NF-kB activity. Site-directed mutagenesis showed that the transfection of the mutated NF-kB binding sites from the PGL3-MUC5AC-3752 promoter luciferase reporter plasmid decreased the luciferase activity after NE stimulation. Among the MAPKs, only extracellular signal-regulated kinases (ERK) were involved in this NE-induced MUC5AC mucin expression. RT-PCR also showed that NE increased MUC5AC mRNA. An EMSA autoradiogram revealed that NE induced NF-kB: DNA binding. CONCLUSIONS: These results indicate that human NE induces MUC5AC mucin through the epidermal growth factor receptor (EGF-R), ERK, and NF-kB pathways in A549 cells.

Paradoxical effects of elastase inhibitor guamerin on the tissue repair of two different wound models: sealed cut and exposed tongue wounds

Innate elastase inhibitors are known to be putatively involved in the regulation of tissue inflammation by inhibiting polymorphonuclear leukocyte (PMN) derived proteinases. The aim of this study was to evaluate affects of leukocyte elastase suppression and PMN infiltration on wound healing in mouse by administering the recombinant elastase inhibitor guamerin (rEIG) in two different wound models; 1) impaired pin-punctured dorsal mucosa of anterior tongue wound, 60 mice, treated with saline containing rEIG that were fed ad libitum and 2) stable linear excisional cutaneous wound, 40 mice, covered with fibrin sealant containing rEIG. The progress of healing was analyzed by histological methods. The tongue wounds treated with rEIG became edematous around the pin-punctured tongue wound, and influx of inflammatory cells and PMN into the underlying stromal tissue were seen rapidly after wounding and peaked between 2-4 days. Whereas the control mice showed almost no wheal formation in the pin-punctured wound, a far lesser levels of PMN infiltration, and almost complete wound closure in 4 days. In the other model, the liner excisional cutaneous wound treated with fibrin sealant containing rEIG showed early wound constriction, lesser degree of inflammatory cells influx, and complete reepithelialization in 4-5 days, whereas the wound of control mice with the fibrin sealant alone showed contrary delayed reepithelialization, greater degree of inflammatory cell infiltration, and consequencial formation of greater granulation tissue at wound site. Taken together, these data suggest paradoxical effects of rEIG on the wound healing where in the wound exposed to infiltrating milieu of microorganisms in the oral cavity, the rEIG aggravates the wound healing by interfering with other innate defensive factors and extended greater flux of PMNs to inflamed wound site, while in the wound enclosed by fibrin, the rEIG accelerated wound healing by inhibiting the inflammation-generated proteases and the acute inflammatory reaction.

Changes in protease in ischemia-reperfusion lung injury and the protective effect of protease inhibitor in rabbit / 解放军医学杂志

Objective To study the changes in protease and cytokines,and to evaluate the protective effect of ulinastatin on ischemia-reperfusion lung injury in rabbits.Methods Thirty New Zealand rabbits were randomly divided into three groups(n=10): control group,traumatic hemorrhagic shock group(shock group) and ulinastatin-treated group.Blood samples were taken at four time points: pre-shock,post-shock,as well as 2h and 4h after blood volume repleshment.The serum levels of TNF-? and IL-8 were measured.4h after blood volume repleshment,the rabbits were sacrificed.The activity of myeloperoxidase(MPO) in lung tissue and neutrophil elastase(NE) in bronchoalveolar lavage fluid(BALF) were determined.Results In control group,the serum concentrations of TNF-? and IL-8 showed no obvious changes during the experiment.While in shock group and ulinastatin-treated group,the serum concentrations of TNF-? and IL-8 increased significantly in the duration of experiment compared with the values before shock(P