Grape Seed Extract Attenuates Demyelination in Experimental Autoimmune Encephalomyelitis Mice by Inhibiting Inflammatory Response of Immune Cells / 中国结合医学杂志

OBJECTIVE@#To examine the anti-inflammatory effect of grape seed extract (GSE) in animal and cellular models and explore its mechanism of action.@*METHODS@#This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis (MS)-experimental autoimmune encephalomyelitis (EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17 and the inflammatory factors such as tumor necrosis factor- α (TNF- α), interleukin-1 β (IL-1 β), IL-6, IL-12, IL-17 and interferon-γ (IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay (ELISA), immunofluorescence staining and Western blot, respectively.@*RESULTS@#GSE reduced the secretion of TNF-α, IL-1 β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide (P<0.01), inhibited the secretion of TNF-α, IL-1 β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days (P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors (P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4 (TLR4) and Rho-associated kinase (ROCKII, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors (P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45+CD11b+ and CD45+CD4+ cells, and weakened the differentiation of Th1 and Th17 (P<0.05). Moreover, it reduced the secretion of inflammatory factors (P<0.01), and prevented the activation of microglia (P<0.05).@*CONCLUSION@#GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.

Application progress on contrast-enhanced ultrasound in acute rejection after kidney transplantation / 器官移植

Early diagnosis of acute rejection is of significance for the protection of renal allograft function. Pathological puncture biopsy is the gold standard for the diagnosis of acute rejection of renal allografts. Nevertheless, it may provoke multiple complications, such as bleeding, infection and renal parenchymal injury, which limit its widespread application. In recent years, the sensitivity of contrast-enhanced ultrasound in the diagnosis of acute rejection has been constantly improved. Ultrasound-targeted microbubble technique has further enhanced the diagnostic specificity of contrast-enhanced ultrasound, making it possible to replace pathological puncture biopsy. Besides, in the field of acute rejection treatment, microbubble ultrasonic cavitation may promote local delivery of immunosuppressants by inducing sonoporation and exhibit anti-rejection effect. In this article, the application of contrast-enhanced ultrasound in the diagnosis and treatment of acute rejection after kidney transplantation was reviewed, aiming to provide reference for widespread application of contrast-enhanced ultrasound in kidney transplantation.

Research progress in involvement of inflammatory cells in formation of coronary collateral circulation / 上海交通大学学报（医学版）

There are abundant anastomotic branches among the branches of coronary artery. When the main coronary artery is occluded, the coronary collateral circulation is formed, which increases the local perfusion of ischemic myocardium and reduces the area of myocardial infarction. There are two main forms of coronary collateral circulation: arteriogenesis and angiogenesis. When coronary artery occlusion occurs, different types of inflammatory cells are recruited into the collateral circulation formation area, which plays an important role in coronary collateral circulation formation. This article discusses the effects of different types of inflammatory cells on the formation of coronary collateral circulation, and further describes the contributory factors of poor coronary collateral circulation formation in diabetic patients.

Phenotypic Characteristics and Differentially Expressed Genes Enrichment Pathway of Inflammatory Cells in Gastritis With Cancerous Potential / 胃肠病学

Background: Gastric cancer, especially intestinal-type gastric cancer, is considered as the result of the process of non-atrophic gastritis-atrophic gastritis-dysplasia-carcinogenesis. Aims: To explore the phenotypic characteristics and differentially expressed genes (DEGs) enrichment pathway of inflammatory cells between gastritis that prone to canceration (atrophic gastritis) and gastritis that not prone to canceration (non-atrophic gastritis). Methods: The datasets of GSE2669, GSE83389, GSE106656 and GSE27411 were downloaded from GEO database. DEGs were screened by R language and verified by GSE116312 dataset. DEGs screened from 3 datasets of 'non-atrophic gastritis-atrophic gastritis' were overlapped with inflammatory cell phenotypic characteristic genes. REACTOME and KEGG analyses of DEGs were performed. Results: A variety of DEGs in the 'normal gastric mucosa-non-atrophic gastritis-atrophic gastritis' dynamic process were screened, and 5 common genes were verified by GSE116312 dataset. A total of 85 inflammatory cell phenotypic characteristic genes were screened from 3 datasets. The percentage of neutrophil was high during 'normal gastric mucosa-non-atrophic gastritis' process while the percentages of fibroblast and macrophage were high during 'non-atrophic gastritis-atrophic gastritis' process. REACTOME and KEGG analyses showed that DEGs of inflammatory cell phenotype during 'non-atrophic gastritis-atrophic gastritis' process were mainly enriched in IL-10, IL-4 and IL-13 signaling pathways and antigen presentation pathway. Conclusions: Macrophage, neutrophil and fibroblast are the inflammatory cell phenotypic characteristics of gastritis with cancerous potential, which enriched in IL and antigen presentation pathways.

Mechanism of effect of IL-17C on survival of kidney graft in mice / 器官移植

Objective To investigate the effect and mechanism of interleukin (IL)-17C in mice undergoing kidney transplantation. Methods The life-supporting kidney transplantation mice models were established using Balb/c (H-2Kd) mice as the donors, IL-17C gene knock out (IL-17CKO) mice (knockout group) and C57BL/6J(H-2Kb) mice (wild group) were chosen as the recipients. The postoperative body mass and survival time of mice were statistically compared between two groups. Pathological examination of the kidney graft was performed by using hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining. The expression levels of granzyme B, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-6 and IL-1β messenger ribonucleic acid (mRNA) in the kidney graft tissue were quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). The proportion of inflammatory cell infiltration in the kidney graft tissue was detected by flow cytometry. Results In the knockout group, the survival time of mice after kidney transplantation was significantly shorter than that of the wild mice (P=0.031). The body mass was more evidently decreased in the knockout group with no statistical significance from that in the wild group. Pathological examination demonstrated that the kidney graft injury in the knockout group was significantly worse than that in the wild group. The mRNA expression levels of granzyme B, IFN-γ, TNF-α, IL-6 mRNA in the knockout group were significantly up-regulated compared with those in the wild group (all P < 0.01). The mRNA expression level of IL-1β showed a decreasing trend with no statistical significance (P=0.16). Flow cytometry analysis revealed that the infiltration of CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6Chi monocyte in the kidney graft of knockout mice was significantly higher compared with that of the wild mice (P < 0.05, P < 0.01), whereas the infiltration of CD45+Ly6ChiF4/80+ macrophage did not significantly differ between two groups (P > 0.05). Conclusions IL-17C participates in the regulation of inflammatory response after kidney transplantation. It can alleviate acute rejection and improve the survival of kidney graft by down-regulating the expression of pro-inflammatory cytokines and infiltration of inflammatory cells.

Predictivity of inflammatory cell counts in early diagnosis of hydatidiform mole

Background: Hydatidiform mole (HM) is the most frequently encountered disease among gestational trophoblastic diseases. HM can invade myometrium and result in hysterectomy and because of the absence of any predictive method, the disease  can  be lately diagnosed in the periphery. Author aimed to evaluate predictive value of  the inflammatory cell counts in molar pregnancies in this study.Methods: Nineteen (19) cases with histopathologic HM diagnosis and 19 cases of control group with pregnancy termination or abortion material reached to a university hospital's pathology department on the same day were included in the study. The data on the same day or the day before the operation was used as the hemogram data.Results: The mean age of the cases were 33.84±8.477. The mean of neutrophil, lymphocyte, monocyte, basophil and eosinophil numbers of the HM group and control group were compared in the 95% confidence interval with the independent t test. No statistical significance was observed in any of the inflammatory cell means  (p>0.05). The ratio of lymphocyte means was statistically significant (p=0.006).Conclusions: In this study, author assessed whether the inflammatory cell counts were a predictive in detecting HM. The statistically significant results that author founded in the means of lymphocyte, suggests that this finding may be predictive of early diagnosis. They concluded that this result can be routinely used after the confirmation of the results in larger series of cases.

Practice experience of establishment of abdominal heart transplantation model combined with tail vein injection in mice (with video demonstration) / 器官移植

Objective To summarize the practice experience of establishing a stable abdominal heart transplantation model combined with tail vein injection in mice. Methods In the preliminary experiment, 50 pairs of donor and recipient Kunming mice received isotransplantation, 40 pairs of donor and recipient C57BL/6J mice underwent isotransplantation. In the formal experiment, 10 pairs of donor and recipient C57BL/6J mice received isotransplantation, 30 pairs of Balb/c mice as the donor and C57BL/6J mice as the recipient received allotransplantation. The time of each step of the heart transplantation (including harvesting and dressing of the donor heart, vascular anastomosis of the recipient, etc.) was recorded. The duration of transplanted heart beat and the survival time of the recipient was observed daily after operation. The time required for tail vein injection in the transplanted mice was recorded. Pathological examination of the transplanted heart was performed at 30 d after isotransplantation (n=5) and 7 d after allotransplantation (n=5). Results In the formal experiment, the success rate of heart transplantation was 90%. The harvesting and dressing time of donor heart was (13.9±0.6) min. The cold ischemia time of the recipient was (14.2±1.2) min. The vascular anastomosis time was (34.2±3.1) min. The total operation time was (86.6±5.4) min. Postoperatively, the transplanted heart of the mice undergoing isotransplantation survived longer than 100 d. Pathological examination at postoperative 30 d demonstrated only a slight amount of inflammatory cell infiltration. The survival time of the mice receiving allotransplantation was (7.2±0.5) d due to rejection reaction. At postoperative 7 d, pathological examination showed a large quantity of inflammatory cells infiltrating into the myocardium, manifested with acute cellular rejection. The success rate reached 90% after over 200 times of tail vein injection. Conclusions In this study, a stable mouse abdominal heart transplantation model is successfully established. The mouse models in the preliminary experiment can be utilized for tail vein injection.

Effect of protein kinase C β inhibitor on renal ischemia-reperfusion injury and expression level of macrophage subtypes in rat models / 器官移植

Objective To investigate the effect of protein kinase C (PKC) β inhibitor on the renal ischemia-reperfusion injury (RIRI) in rat models and detect the expression level of macrophage subtypes. Methods Eighteen healthy SD male rats were randomly divided into the Sham operation group (Sham group, n=6), RIRI group (n=6), PKCβ inhibitor +RIRI group (Inhibitor+RIRI group, n=6). Serum and left renal tissue samples were collected at postoperative 24 h. The contents of serum creatinine (Scr) and blood urea nitrogen (BUN) were detected by automatic biochemical analyzer. The infiltration of inflammatory cells and pathological injury in the renal tissues were observed by hematoxylin-eosin (HE) staining. The expression levels of CD68, inducible nitric oxide synthase (iNOS) and CD206 proteins in the renal tissues of rats in each group were assessed by immunohistochemistry and Western blot. Results The contents of serum Scr and BUN in the RIRI group were significantly higher than those in the Sham group (both P < 0.05). The contents of serum Scr and BUN in the Inhibitor+RIRI group were considerably lower than those in the RIRI group (both P < 0.05). No obvious renal injury was noted in the Sham group, whereas renal inflammatory cell infiltration and renal tubular structural damage were observed in the RIRI group. The renal inflammatory cell infiltration and renal tubular structural damage in the Inhibitor+RIRI group was slighter than that in the RIRI group. The protein expression levels of CD68, iNOS and CD206 in the renal tissue of rats in the RIRI group were significantly higher than those in the Sham group (all P < 0.05). The protein expression levels of CD68 and iNOS in the Inhibitor+RIRI group were remarkably lower than those in the RIRI group (all P < 0.05). The expression level of CD206 protein in the Inhibitor+RIRI group was significantly higher than that in the RIRI group (P < 0.05). Conclusions PKC β inhibitor can alleviate the RIRI in rat models to certain extent, which may be correlated with the role of PKC β inhibitor in mitigating inflammatory cell infiltration in ischemic renal tissues and promoting the expression of alternatively activated macrophage

An Important Role of Macrophages for Wound Margin Regeneration in a Murine Flap Model

BACKGROUND: Macrophages have been known to have diverse roles either after tissue damage or during the wound healing process; however, their roles in flap wound healing are poorly understood. In this study, we aimed to evaluate how macrophages contribute to the flap wound regeneration.METHODS: A murine model of a pedicled flap was generated, and the time-course of the wound healing process was determined. Especially, the interface between the flap and the residual tissue was histopathologically evaluated. Using clodronate liposome, a macrophage-depleting agent, the functional role of macrophages in flap wound healing was investigated. Coculture of human keratinocyte cell line HaCaT and monocytic cell line THP-1 was performed to unveil relationship between the two cell types.RESULTS: Macrophage depletion significantly impaired flap wound healing process showing increased necrotic area after clodronate liposome administration. Interestingly, microscopic evaluation revealed that epithelial remodeling between the flap tissue and residual normal tissue did not occurred under the lack of macrophage infiltration. Coculture and scratch wound healing assays indicated that macrophages significantly affected the migration of keratinocytes.CONCLUSION: Macrophages play a critical role in the flap wound regeneration. Especially, epithelial remodeling at the flap margin is dependent on proper macrophage infiltration. These results implicate to support the cellular mechanisms of impaired flap wound healing.

Effects of FEA from Polygonum hydropiper L. on Biochemical Indexes and Inflammatory Cytokines in Mice with Endotoxemia Induced by Lipopolysaccharide / 中国中医药信息杂志

Objective To observe the effect of flavonoids ethyl acetate(FEA)from Polygonum hydropiper L.on biochemical indexes and inflammatory cytokines in mice with endotoxemia; To expore the mechanism. Methods Total flavonoids in the whole plant of Polygonum hydropiperum L. were extracted by enzymolysis-ultrasonic coupling method. The FEA part were obtained by extracting and separating, followed with macroporous resin purification and enrichment. The animal model of endotoxemia was established by stimulation of lipopolysaccharide (LPS) in mice. Experimental mice were divided into blank group, model group, FEA high-, medium-, and low-dose groups. Each administration group was given the corresponding concentration of herb liquor. The concentration of malondialdehyde (MDA) and myeloperoxidase (MPO) in intestinal tissue, total antioxidant capacity (T-AOC), superoxide dismutase (T-SOD) activity and glutathione peroxidase (GSH-Px) activity in liver tissue, glutathione (GSH), lysozyme (LZM) and acid phosphatase (ACP) in serum were determined. The contents of tumor necrosis factor-α (TNF-α), interferon (IFN)-α, IFN-γ and interleukin-2 (IL-2) in lung tissue were detected by RT-PCR. Results Compared with blank group,the levels of MDA, MPO in intestinal tissue and serum ACP of model mice were increased, while T-AOC, T-SOD, GSH-Px in liver tissue, serum GSH and LZM levels were decreased; TNF-α in serum, intestinal and liver tissues were increased, the expressions of TNF-α, IFN-α, IFN-γ and IL-2 mRNA in lung tissue were increased. Compared with the model group, the levels of MDA, MPO in intestinal tissue and serum ACP were decreased in all dose of FEA groups;The levels of T-AOC, T-SOD, GSH-Px in liver tissue, serum GSH and LZM of FEA medium and high-dose groups were increased. The content of TNF-α in mice serum, intestinal and liver tissues of all dose of FEA groups were significantly reduced, and the expressions of TNF-α, IFN-α, IFN-γ and IL-2 mRNA in lung tissues were significantly decreased. The pathological morphology of mice lung, ileum and colon tissues of FEA high-dose group were significantly ameliorated than model group. Conclusion FEA can attenuate inflammation injury of endotoxemia mice induced by LPS, which has protective effects for organism.

The transition of sputum inflammatory cell profiles is variable in stable asthma patients

BACKGROUND: The sputum inflammatory cell profile is an important indicator for classifying asthma phenotypes. OBJECTIVE: To investigate if sputum inflammatory cell profile remains stable and there are different characteristics between groups that show different profile over time in stable asthmatic patients. METHODS: A total of 149 asthmatic patients, who were clinically stable at the time of sputum examination and had undergone sputum analysis twice, were subjected to a detailed review. Eosinophilic inflammation was diagnosed when the proportion of the sputum eosinophils was >3%. We divided the patients into 4 groups according to the transition patterns of their sputum profiles: group 1, persistent eosinophilia; group 2, eosinophilic to noneosinophilic; group 3, noneosinophilic to eosinophilic; and group 4, persistent noneosinophilia. The results of the pulmonary function tests and other clinical parameters were compared between these 4 groups. RESULTS: Thirty-four of the initially eosinophilic asthmatic patients (39.5%; 34 of 86 patients) demonstrated noneosinophilic airway inflammation at their second sputum examination, and 24 of the initially noneosinophilic patients (38.1%; 24 of 63 patients) demonstrated eosinophilic airway inflammation at follow-up. Various clinical parameters, except the blood eosinophil count, demonstrated no significant differences between the eosinophilic and noneosinophilic asthmatic patients or among the 4 groups. CONCLUSION: A substantial proportion of asthmatic patients who demonstrate a certain sputum inflammatory cell profile at the initial examination demonstrated profile transition in clinically stable settings over time. The clinical significance of using induced sputum analysis to phenotype stable asthmatic patients requires further evaluation.

Effect of Oxymatrine Ointment on Mice with Chronic Eczema / 中国药师

Objective: To observe the effect of oxymatrine ointment on chronic eczema in mice, and preliminarily explore the mechanism. Methods:Thirty-two Kunming mice were randomly divided into four groups including oxymatrine ointment group, blank model group, blank ointment group and positive drug group treated with compound dexamethasone acetate cream. Eczema skin was con-tinuously treated with drugs for 14 days. The mice were sacrificed on the 15th day, and the eczema skin was clipped from back to make histological sections. The changes of inflammation were observed, and the inflammatory cell count was obtained. Meanwhile, heart blood was collected, and serum was obtained by centrifugation. The serum levels of IL-4 and IL-1β were determined by ELISA. Re-sults:The inflammatory cell count in oxymatrine ointment group and the positive drug group was lower than that in the blank model group and the blank ointment group (P0. 05). Conclusion: Oxymatrine ointment has certain anti-inflammatory effect on eczema, and the mechanism needs to be studied further.

Expression of inflammatory factors in hippocampal subregions of rats after cerebral ischemia-reperfusion / 军事医学

Objective To detect the damage of hippocampal neurons and the changes in inflammatory cytokines in rats after cerebral ischemia-reperfusion(I/R)and compare the expressions of IL-1β,IL-6 and TNFαin hippocampal DG,CA1 and CA3 subregions.Methods The focal cerebral I/R model was induced by an intraluminal filament embolism.The SD rats were randomly divided into the sham-operated group(SHAM group)and the middle cerebral artery occlusion-reperfusion group(MCAO group).HE staining was employed to detect the damage to hippocampal DG, CA1 and CA3 subregions.The expression levels of IL-1β, IL-6 and TNFα were detected by immunofluorescence assay.Results Compared with SHAM group,hippocampal DG,CA1 and CA3 subregion neurons in MCAO group were severely damaged, with occurred inflammatory cell infiltration,and a large amount of neurons apoptosis, and the expressions of IL-1β, IL-6 and TNFαin each subregion increased significantly.At the same time, in MCAO group, the expression of inflammatory cytokines in CA1 subregion was more significant than that in DG and CA 3 subregions(P<0.05).Conclusion Cerebral I/R could cause neuronal damage, inflammatory cell infiltration, and neuronal apoptosis in the DG, CA1 and CA3 subregions of the hippocampus and increase the release of inflammatory cytokines.In MCAO group, the expression of inflammatory cytokines in CA1 subregion of hippocampus is significantly higher than that in DG and CA 3 subregions, suggesting that CA1 region is more sensitive to I/R injury.

Clinical observation of the efficacy of No. 1 Baiban granules on vitiligo / 天津医药

Objective To observe the efficacy of No. 1 Baiban granules in the treatment of vitiligo using skin confocal laser scanning microscopy. Methods The study was approved by the hospital ethics committee. Nine-two patients with qi stagnation and blood stasis were collected from Department of Dermatology of the Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine and were randomly assigned to two groups. The experimental group (n=49) received oral Baiban granuleΙ, 3 g, twice a day, transfer factor capsules 6 mg, 3 times a day, 0.1%mometasone furoate cream (excluding the face) for external use, once a day, or 0.1%tacrolimus ointment (Afacial), twice a day for 3 months. The control group (n=43) only received oral transfer factor capsules 6 mg, 3 times a day, 0.1%mometasone furoate cream (except for the face) for external use, once a day, or 0.1%tacrolimus ointment (face), twice a day, treatment for 3 months. The therapeutic effect was observed in two groups. CLSM was used to monitor the compound color and pigment recovery level, and changes of the number of dendritic cells and inflammatory cell infiltration in skin. Results The compound color was improved significantly after treatment in experimental group and control group. The total effective rate was higher in experimental group than that of control group (81.63% vs. 69.77%,χ2=3.947,P<0.05). Results of CLSM observation showed that the pigment recovery of skin lesion, the proportion of dendritic cells and inflammatory cells were significantly higher in experimental group than those of control group (P < 0.05). Conclusion The treatment effect of No. 1 Baiban granules is more obvious for vitiligo of qi stagnation and blood stasis.

Induction of long-term heart survival after secondary transplantation by anti-RANTES monoclonal antibody combined with ciclosporin in mouse models / 器官移植

Objective To evaluate the effect of anti-RANTES monoclonal antibody in combination with ciclosporin (CsA)upon inhibiting the rejection response during secondary heart transplantation in mouse models. Methods BALB/c mouse models were used as the donors and C57BL/6 mice were utilized to establish secondary heart transplantation recipient models. The animals were randomly divided into the control (physiological saline,n =6 ),A (anti-RANTES monoclonal antibody treatment,n =6 ),B (CsA treatment,n =6 ) and C groups (anti-RANTES monoclonal antibody combined with CsA treatment,n=6). The survival time of heart after secondary transplantation was observed. The degree of acute heart rejection was assessed by histopathological analysis. The relative expression levels of RANTES,interleukin(IL)-2,IL-1 0,interferon(IFN)-γand transcription growth factor(TGF)-βmessenger ribonucleic acid (mRNA)in the heart grafts were quantitatively measured by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The serum levels of RANTES,IFN-γ,IL-2,IL-1 0 and TGF-βwere detected by enzyme-linked immune absorbent assay (ELISA). Results The heart grafts of all mice survived after secondary cardiac transplantation. Compared with the control group,the survival time of hearts in group A,B and C was significantly prolonged (all P<0. 01 ). Pathological staining revealed that the quantity of infiltrated inflammatory cells in group C was significantly decreased than those in the other groups. The expression levels of heart RANTES,IFN-γand IL-2 mRNA in group C were significantly down-regulated, whereas the expression levels of IL-1 0 and TGF-βmRNA were considerably up-regulated compared with those in the other three groups (all P<0. 05). The serum levels of RANTES,IL-2 and IFN-γin group C were significantly down-regulated, whereas the serum contents of IL-1 0 and TGF-βwere considerably up-regulated compared with those in the other three groups (all P<0. 05 ). Conclusions Combined application of anti-RANTES monoclonal antibody and CsA can effectively induce the immune tolerance to secondary cardiac transplantation and prolong the survival time of the cardiac grafts in mouse models.

Macrophage infiltration in the wound healing of diabetic rats / 医学研究生学报

Objective Knowledge about the infiltration of macrophages in diabetic wounds can help to figure out the pathogene -sis of poor healing of diabetic wounds .The aim of this study was to observe the macrophage infiltration and the expression of relative in-flammatory factors during the wound healing of diabetic rats and explore the relationship between macrophages and diabetic wound heal -ing. Methods Male Sprague Dawley rats were randomly divided into STZ induced diabetic group and normal control group .Each group had 15 rats.A 1 cm2 full-thickness skin defect was created on the rat dorsum .Wound samples and was excised .On post injury day (PID) 3, 7, and 14, rats of each group were sacrificed after wound samples and tissues around the wound edge were obtained .The differences of the wound closure rate , macrophage infiltration and the relative mRNA expression of the inflammatory factors from macro-phages were observed . Results The wound closure rate was lower in diabetic group on PID 3, 7, and 14 ([29.5 ±5.4]%vs [45.9 ± 12.8]%, [71.6 ±3.1]%vs [80.1 ±6.9]%, [93.9 ±2.8]%vs [99.4 ±1.4]%, P<0.05).HE staining showed inflammatory cells infil-diabetic wound tissue on PID 3 (P<0.05) and a higher expression on PID 14 (P<0.05).The CCR7 fluorescence staining showed more positive staining cells stayed in diabetic wound on PID 14. Conclusion Macrophage infiltration decreases in the early phase of diabetic wound healing and sustains in wound tissue in advanced stage accompanied by the expression change of related inflammatory factors, which could contribute to the difficult wound healing of diabetic rats .

Resistance of Berberine on Lung Inflammatory Injury in Mice with Viral Pneumonia Caused by Influenza Virus / 中国中医药信息杂志

Objective To observe the effects of berberine on ICAM-1, VCAM-1 expression and inflammatory cells exudation in mice with viral pneumonia caused by influenza virus, and explore its anti-injury effect. Methods Totally 108 BALB/c mice were randomly divided into control group, model group, and berberine group. 25 μL 50 LD50 influenza virus, mouse lung-adapted strain, was intranasally inoculated to model group and berberine group. 1 h after infection, control and model group were intragastrically given 25 μL distilled water, berberine group was treated by intraperitoneal injection with berberine at a dose of 0.005 g/(kg·d) for 5 days, twice per day. On day 2, 4 and 6 after infection, immunocytochemical method was used to detect ICAM-1 and VCAM-1 expression, and sorting cell count of leukocytes in bronchoalveolar lavage fluid (BALF) were counted. Results The expression of ICAM-1 and VCAM-1 in model group increased obviously on day 2, 4, 6, and which in berberine group decreased compared with model group (P<0.01). WBC, mononuclear cell, eosinophile cell and neutrophil cell number in model group increased significantly. WBC and neutrophil cell number decreased in berberine group on day 6 (P<0.01), and the mononuclear cell number decreased on day 4 (P<0.01). Conclusion Berberine inhibited the expression of ICAM-1 and VCAM-1, and decreased the inflammatory cells exudation in lung of mice with viral pneumonia caused by influenza virus. Berberine has protective effect on inflammatory injury of lung tissue in mice with viral pneumonia caused by influenza virus.

A comparative study of apoptosis in reticular and erosive oral lichen planus

The oral lichen planus (OLP) is a chronic inflammatory disease, probably autoimmune, with different clinical forms. The most common types are the reticular and the erosive ones. Apoptosis participates in the destruction of basal keratinocytes, but its role in the perpetuation of the subepithelial lymphocytic infiltrates was not yet investigated. To evaluate the involvement of apoptosis in the epithelium and in subepithelial lymphocytic infiltrates, 15 samples of reticular and erosive OLP and 10 samples of healthy oral mucosa were collected and processed histologically. Apoptosis was quantified in the epithelium and in inflammatory cell infiltrates. TUNEL reaction was used to measure apoptosis in the infiltrates. Erosive OLP showed more intense epithelial apoptosis than reticular OLP and controls. In contrast, apoptosis in the inflammatory cell infiltrates was more frequent in reticular than in erosive OLP. Lymphocytes were the predominant cells within the inflammatory cell infiltrates and were more frequent in erosive OLP than in reticular type. These results suggest that different apoptotic levels are involved in the erosive/reticular switch in OLP, determining different clinical presentations. In conclusion, decreased apoptosis in inflammatory infiltrates may contribute to the persistence of T lymphocytes, worsening the attack to the epithelium in erosive OLP.

O líquen plano oral (LPO) é uma doença crônica inflamatória, provavelmente auto-imune, com diferentes formas clínicas. Os tipos mais comuns são o reticular e o erosivo. A apoptose participa da destruição dos ceratinócitos basais, no entanto o seu papel na perpetuação do infiltrado linfocitário subepitelial ainda não foi investigado. Para avaliar o envolvimento da apoptose no epitélio e no infiltrado linfocitário subepitelial, quinze amostras de LPO reticular, quinze de LPO erosivo e dez amostras de mucosa oral saudável foram coletadas e processadas histologicamente. A apoptose foi quantificada no epitélio e nas células do infiltrado inflamatório. A reação de TUNEL foi usada para mensurar a apoptose no infiltrado. A intensidade da apoptose no epitélio mostrou ser maior no LPO erosivo que no LPO reticular e estes foram maiores que no controle. Em contraste, a apoptose nas células do infiltrado inflamatório foi mais freqüente no LPO reticular que no LPO erosivo. Os linfócitos foram as células predominantes dentro do infiltrado inflamatório e foram mais freqüentes no tipo erosivo de LPO que no tipo reticular. Estes resultados sugerem que diferentes níveis de apoptose estão envolvidos no tipo erosivo e reticular de LPO, determinando as diferenças nas apresentações clínicas. Em conclusão, a diminuição da apoptose no infiltrado inflamatório pode contribuir para a persistência dos linfócitos T, piorando o ataque ao epitélio no LPO erosivo.

Immunologic Evaluation of Drug Allergy

Hypersensitivity drug reactions (HDR) consist of an individual abnormal response with the involvement of the immunological system. In addition to specific immunological mechanisms where specific antibodies or sensitised T cells participate, release of inflammatory mediators by non-specific immunological recognition may also occur. Within this category are one of the most common groups of drugs, the non-steroidal anti-inflammatory drugs. In addition to chemical drugs new emerging ones with an increasing protagonism are biological agents like humanised antibodies and others. For IgE dependent reactions both in vivo and in vitro tests can be used for the immunological evaluation. Sensitivity of these is not optimal and very often a drug provocation test must be considered for knowing the mechanism involved and/or establishing the diagnosis. For non-immediate reactions also both in vivo and in vitro tests can be used. Sensitivity for in vivo tests is generally low and in vitro tests may be needed for the immunological evaluation. Immunohistochemical studies of the affected tissue enable a more precise classification of non-immediate reactions. The monitorization of the acute response of the reactions has given clues for understanding these reactions and has promising results for the future of the immunological evaluation of HDR.

Diltiazem inhibits cytokine expression in rat myocardium of calcium paradox model / 中国药理学通报

Aim To explore the cardiac cytokine expression in rat model of myocardial calcium overload, and the intervention from diltiazem.Methods The intracellular Ca~(2+) overload was induced by the isolated rat heart subjected to 5 min Ca~(2+) depletion and 30 min Ca~(2+) repletion (Ca~(2+) paradox) by the Langendorff technique.There were five groups in this study, including Ca~(2+) overload group, normal control group, Ca~(2+) depletion control group, Ca~(2+) overload-diltiazem group, and Ca~(2+) depletion-diltiazem group.The views of myocardial pathology and ultrastruction were observed by electron microscope and light microscope respectively. The cardiac intracellular [Ca~(2+)]_i was detected by atom spectrophotometer. The expression of TNF-α, IL-1β, L-6, TGF-β1, and IL-10 was detected by RT-PCR method.Results In Ca~(2+) overload group, few inflammatory cells were found in myocardium under the light microscope. And the views of electron microscope presented that cardiocyte membranes, nucleolus, and mitochondria were disorganized obviously.Compared with normal control group, the inflammatory cytokines as TNF-α, IL-1β, and IL-6 were increased significantly whereas there was nearly no difference of the expression of TGF-β1 and IL-10 in Ca~(2+) overload group.Ca~(2+) overload-diltiazem group showed that TNF-α, IL-1β, and IL-6 were decreased significantly. There were no statistical differences in the structure of myocardium, intracellular [Ca~(2+)]_i, and cardiac cytokines expressions in the three control groups, including normal control group, Ca~(2+) depletion control group and Ca~(2+) depletion-diltiazem group.Conclusions Instead of TGF-β1 and IL-10, the expression of TNF-α, IL-1β, and IL-6 is increased obviously in myocardium of calcium paradox model. Diltiazem can inhibit the cardiac expression of TNF-α, IL-1β, and IL-6 induced by myocardial calcium overload.