Mastócitos e síndrome de ativação mastocitária: o que há de novo? / Mast cells and mast cell activation syndrome: what's new?

Os mastócitos são as principais células efetoras da resposta alérgica aguda, desempenhando também um papel importante na angiogênese, tolerância imunológica, regulação da fibrinólise, regeneração neuronal e osteoclastogênese. Localizam-se maioritariamente na pele e nas mucosas do intestino e pulmões, onde exercem uma função "sentinela". As síndromes de ativação mastocitária são caracterizadas pela ocorrência de episódios recorrentes de manifestações clínicas resultantes da libertação de mediadores mastocitários. Esta constitui-se como entidade complexa com um espectro de sintomas associados, representando um desafio diagnóstico e terapêutico. Nesta revisão, os autores pretendem apresentar uma visão geral sobre a estrutura e função dos mastócitos e sobre os critérios diagnósticos e abordagem terapêutica da síndrome de ativação mastocitária.

Mast cells are the main effector cells of acute allergic response, also playing an important role in angiogenesis, immune tolerance, regulation of fibrinolysis, neuronal regeneration, and osteoclastogenesis. They are generally located in the skin and mucous membranes of the intestines and lungs, where they perform a "sentinel" function. Mast cell activation syndrome is characterized by recurrent clinical manifestations resulting from the release of mast cell mediators. This complex entity, which involves a spectrum of associated symptoms, is a diagnostic and therapeutic challenge. In this article we overview of the structure and function of mast cells, in addition to the diagnostic criteria and therapeutic approaches to mast cell activation syndrome.

Effects and mechanism of anaphylactoid reaction induced by nonapeptide activating mast cells / 中国药房

OBJECTIVE To study the effects and potential mechanism of anaphylactoid reaction induced by nonapeptide IVQKIKHCF activating mast cells. METHODS Using human mast cell line LAD2 as subject， and substance P as positive control， the activation effects of 25， 50 and 100 μmol/L IVQKIKHCF on mast cells were investigated by determining the release rate of β-aminohexosidase， histamine release， and the contents of inflammatory factors； using MrgprX2-knockdown LAD2 cells and Mas- related G protein-coupled receptor X2 （MRGPRX2） high-expression human embryonic kidney cell line HEK293 （MRGPRX2/ HEK293 cells） as subject， the correlation between the activation effect of IVQKIKHCF and MRGPRX2 was investigated by determining the release rate of β-aminohexosidase， and intracellular calcium ion concentration. RESULTS IVQKIKHCF with 25， 50， 100 μmol/L could significantly increase the release rate of β-aminohexosidase and histamine release in LAD2 cells （P＜0.05）， and promote the release of tumor necrosis factor-α， interleukin-8， macrophage inflammatory protein-1β and monocyte chemotactic protein-1 to varying degrees （P＜0.05）. After knocking down MrgprX2， the effects of 25， 50， 100 μmol/L IVQKIKHCF promoting the release of β-aminohexosidase in LAD2 cells were reversed significantly （P＜0.05）， resulting in an increase of calcium ion concentration in MRGPRX2/HEK293 cells. CONCLUSIONS Nonapeptide IVQKIKHCF can promote mast cells to release granular matter and inflammatory mediators by activating MRGPRX2 thus inducing anaphylactoid reaction.

Prognostic significance of mast cells and vascular density in prostatic adenocarcinoma

Context: Though mast cells infiltrate solid tumors, the exact role of mast cells in tumor biology is controversial. Mast cell density (MCD) may vary depending on its location in the tumor and tumor vascularity. MCD may influence the tumor aggressiveness. Aims: This study evaluates MCD and tumor vascularity in different histopathological grades of adenocarcinoma prostate. Settings and Design: Descriptive study with purposive sampling. Methods and Material: The subjects of study were 42 adenocarcinoma patients. 20 cases were of intermediate grade (Gleason score 2–7) and 22 were of high-grade (Gleason score 8-10). Histological diagnosis was made by examining sections stained with hematoxylin and eosin. Additional sections from the same block were stained for mast cells using Giemsa stains as per standard protocol. Mast cell count was done in minimum six random high-power microscopy fields in four different regions- intratumoral, peritumoral, stromal and perivascular regions. Statistical Analysis Used: SSPS software version 13.0. Descriptive statistics, Student's t test and ANOVA test. Results: In high-grade adenocarcinoma, mast cell counts were higher in perilesional, stromal and perivascular regions, whereas it was lower in intralesional areas as compared to the intermediate grade. However, statistical significance was observed only for the perivascular region. There was significantly higher number of blood vessels in high-grade adenocarcinoma as compared to intermediate grade adenocarcinoma. Conclusions: In this study, perilesional mast cells and vascularity increased with increased severity of adenocarcinoma. These findings suggest a possible influence of mast cells on the tumor microenvironment such as vessel density and aggressiveness of tumor. However, further studies are required to substantiate results of this study.

Mastocitosis del adulto: reporte de un caso / Adult mastocytosis: a case report

RESUMEN La mastocitosis es una enfermedad heterogénea, caracterizada por una acumulación de mastocitos en uno o más órganos, siendo el más afectado la piel. Es más frecuente en niños, pero también se presentan casos en los adultos. Hay diferencias significativas entre las formas de presentación en estos grupos etarios, así como también en su evolución y pronóstico. Presentamos el caso clínico de una paciente con mastocitosis cutánea de inicio en la adultez.

ABSTRACT Mastocytosis is a heterogeneous disease, characterized by an accumulation of mast cells in one or more organs. The skin being is the most frecuently affected organ. It is more common in children, but cases also occur in adults. There are significant differences between the forms of presentation in these age groups, as well as in their evolution and prognosis. We report the case of a patient with adult-onset cutaneous mastocytosis.

Prognostic Significance of Lymphatic Vessel Density by D2-40 Immune Marker and Mast Cell Density in Invasive Breast Cancer: A Cross Sectional Study at Tertiary Care Hospital in South India.

Background: Tumour induced lymphangiogenesis plays a crucial role in metastasis and tumour progression. The intratumoural and peritumoral lymphatics are supposed to have different biological effects. The aim of present study was to investigate the correlation of intratumoral lymphatic vessel density (I LVD), peritumoral lymphatic vessel density (P LVD), intratumoral mast cell density (I MCD) and peritumoral mast cell density (P MCD) with prognostic parameters in primary breast carcinoma. Methods: Lymphangiogenesis was detected using D2-40 monoclonal antibody and mast cell by using toludine blue stain in 50 cases of primary breast carcinoma. Positively stained lymphatic vessels were counted at 40 x in dense lymphatic vascular foci (hot-spot) within the tumour. Chi square, ANOVA test and Pearsons correlation was applied to determine the relationship amongst various variables, with statistical significance set at p <0.05. Results: Mean P LVD was significantly higher than I LVD (6.25±21 vs 2.75±2.27,p <0.005). Significant correlation was noted between I LVD and P LVD and age, tumour laterality, tumour size and overall staging. However, there was no correlation between I LVD and P LVD with other important clinicopathologic prognostic markers like grade, lymphnode status and lymphovascular invasion. MCD was higher in both intratumoral and peritumoral location as compared to normal tissue. There was an association noted between P MCD with pathological staging and perineural invasion. However, there was no significant association of I MCD and P MCD with other prognostic markers like grade and lymphnode status. No significant correlation was noted between I LVD, P LVD, I MCD and P MCD. Conclusion: The evidences from our study support the utility of D2-40 stain in determining the lymphatic density in IBC. The study findings also establish the existence of lymphangiogenesis in both intratumoral and peritumoral location. For now, the data presented herein do not permit us to promote the utility of LVD and MCD as predictors of prognosis in invasive breast carcinoma.

Effect of Da Jianzhongtang on Visceral Pain in Rats with Irritable Bowel Syndrome / 中国实验方剂学杂志

Objective:To investigate the effects of Da Jianzhongtang on substance P （SP）， mast cells （MC）， Toll like receptor 2 （TLR2）， TLR4 on MC model and nuclear transcription factor （NF）-<italic>κ</italic>B p65 in visceral pain rats with irritable bowel syndrome （IBS）， and explore its mechanism of action on IBS visceral pain. Method:Forty-eight 3-day-old SD rats were randomly divided into 6 groups： the control group （control）， irritable bowel syndrome group （IBS）， ketotifen group （Ketotifen，0.18 mg·kg^-1）， Da Jianzhongtang low， medium and high dose groups （DJZT-L， DJZT-M， DJZT-H，2.16，1.08，0.54 g·kg^-1）， with 8 rats in each group. Intragastric administration lasted for 2 weeks. Maternal separation method was used to establish the IBS visceral pain model in rats. The visceral sensitivity of rats was evaluated at 60， 40 and 20 mmHg （1 mmHg≈0.133 kPa） with Abdominal wall withdrawal response （AWR） scale. SP and NF-<italic>κ</italic>B p65 protein expression levels in colon tissue were detected with Western blotting technique. TLR2 and TLR4 proteins on mast cell membrane were detected by immunofluorescence staining. The degranulation rate of mast cells in colon tissue was detected by toluidine blue staining. Result:Compared with normal rats， AWR scores of model rats significantly increased at 60， 40， and 20 mmHg pressure （<italic>P</italic><0.05，<italic>P</italic><0.01）， the degranulation rate of mast cells in colon tissue and SP protein expression in colon tissue significantly increased （<italic>P</italic><0.01）， TLR2， TLR4， and nuclear NF-<italic>κ</italic>B p65 expression on mast cell membrane significantly increased （<italic>P</italic><0.01）. Compared with model rats， the AWR scores of DJZT-H group （pressure of 40， 20 mmHg） and DJZT-M group （pressure of 60， 40， 20 mmHg） significantly decreased. Meanwhile， the degranulation rate of colon mast cells， and the SP， TLR2， TLR4， and NF-<italic>κ</italic>B p65 expression also significantly decreased （<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:Da Jianzhongtang can affect mast cell activity and finally decrease visceral pain of IBS rats by down-regulating SP in colon tissue.

Glaucocalyxin A attenuates allergic responses by inhibiting mast cell degranulation through HMGB1/TLR4/NF-κB signaling pathways / 药学学报

The study is to investigate the effect of glaucocalyxin A (GLA) on mast cell-mediated anaphylaxis. The animal welfare and experimental process of this experiment followed the regulations of the Animal Ethics Committee of Yanbian University. BALB/c mice were used in the animal experiment and randomly divided into five groups, control group, model group, and GLA low, medium, and high dose groups (10, 20, and 40 mg·kg-1). Mice were sensitized by intradermal injection of anti-dinitrophenyl-immunoglobulin E (DNP-IgE) into the ears and challenged with a mixture of DNP-human serum albumin (HSA) and 4% evans blue into the tail veins to prepare an animal skin passive cutaneous anaphylaxis (PCA) model, which was collected from both ears for measurement of dye staining and histology. Rat peritoneal mast cells (RPMCs) were used in the cell experiment and divided into control, IgE + antigen (Ag), and IgE + Ag + GLA groups to determine histamine release as well as calcium influx levels. High-affinity IgE receptor (FcεRI)-mediated signaling pathway proteins and HMGB1/TLR4/NF-κB (high mobility group box 1/toll like receptor 4/nuclear transcription factor kappa B) signaling proteins were detected by Western blot. The results of animal experiments suggest that GLA inhibits PCA, reduces evans blue dye exudation, and reduces ear inflammation and ear thickness in mice. The results of cellular experiments suggested that GLA could reduce histamine release and calcium influx, and inhibit tumor necrosis factor-α (TNF-α), interleukin (IL)-4, IL-13, and IL-1β production; Western blot results showed that GLA inhibited FcεRI-mediated phosphorylation levels of spleen tyrosine kinase (Syk), Lck/Yes novel tyrosine kinase (Lyn), tyrosine kinase Fyn (Fyn), growth-factor receptor-bound protein 2 (Gab2), and phospholipase C (PLC) γ1, while GLA inhibited HMGB1/TLR4 signaling pathway to limit NF-κB p65 nuclear metastasis. The results indicate that GLA inhibits mast cell degranulation and attenuates allergic inflammation through the HMGB1/TLR4/NF-κB signaling pathway.

In the search of mobile target for acupuncture: why is mast cell? / 中国针灸

Both clinical practice and basic research of acupuncture have pointed out that acupuncture treatment has specific tissue and cellular targets. In addition to the known fixed tissue targets such as nerves and blood vessels, the author analyzes the biological characteristics of other skin resident cells in the skin and concludes that cutaneous mast cells are the most suitable candidate for the cellular target of acupuncture. A hypothesis of the bionic acupuncture is proposed to explain the biological principles by which the innate immunity and healing system respond to acupuncture. The distribution of mast cells in the human skin is characterized by "approaching to the terminals and gathering at the orifices", and the cell density is highly correlated with the density of acupoints and the micro-acupuncture systems. These evidences all support that mast cells are the mobile target cells for acupuncture, which can explain some clinical phenomena and principles of acupuncture, and suggest mast cells as one of the tissue markers for acupoints.

Therapeutic Efficacies of Wujiwan at Different Compatibilities in Treatment of Irritable Bowel Syndrome / 中国实验方剂学杂志

Objective:To compare the therapeutic efficacies of Wujiwan at two different compatibilities （No.1 and No.2） on irritable bowel syndrome （IBS） based on neuro-endocrine-immune network， and provide a theoretical basis for the treatment based on syndrome differentiation in traditional Chinese medicine （TCM）. Method:The chronic animal model of IBS with visceral hypersensitivity was established by colon irritation via percutaneous transluminal coronary angioplasty （PTCA） in suckling rats. The animals were randomly divided into a control group， a model group， a dicetel group （0.01 g·kg^-1）， low- （0.335 g·kg^-<bold>¹</bold>）， medium- （0.67 g·kg^-<bold>¹</bold>）， and high-dose （1.34 g·kg^-<bold>¹</bold>） No. 1 Wujiwan groups， and low- （0.385 g·kg^-<bold>¹</bold>）， medium- （0.77 g·kg^-<bold>¹</bold>）， and high-dose （1.54 g·kg^-<bold>¹</bold>） No. 2 Wujiwan groups. The thresholds of abdominal elevation and bow back elevation were evaluated to detect the effect of Wujiwan on intestinal sensitivity of IBS. The density of mast cells （MC） in the colonic tissue of model rats was detected by the modified toluidine blue staining method. The concentrations/positive expression of 5-hydroxytryptamine （5-HT）， substance P （SP）， somatostatin （SS）， and vasoactive intestinal peptide （VIP） in the blood/colon tissue were detected by enzyme-linked immunosorbent assay （ELISA） and immunohistochemistry （IHC） assay. Result:There was no significant difference in body weight among different groups. Compared with the control group， the model group exhibited decreased thresholds of abdominal elevation and bow back elevation （<italic>P<</italic>0.01）， increased density of MCs in the colon tissue （<italic>P<</italic>0.05）， up-regulated levels of 5-HT， SP， and SS in the blood and colon tissue （<italic>P<</italic>0.05， <italic>P<</italic>0.01）， and elevated VIP level in the colon tissue （<italic>P</italic><0.05）. Compared with the model group， Wujiwan at different compatibilities could increase the thresholds of abdominal elevation and bow back elevation （<italic>P</italic><0.01）， diminish the count of MC in the colon tissue （<italic>P</italic><0.05）， and reduce the levels of 5-HT， SP， SS， and VIP （<italic>P</italic><0.05）. As demonstrated by the comparison of No. 1 and No. 2 Wujiwan， No. 1 was superior to No. 2 in reducing the concentrations of 5-HT， SP， and SS in the blood， especially in 5-HT （<italic>P</italic><0.01）. No significant difference between No. 1 and No. 2 in reducing 5-HT positive expression in the colon tissue was observed. Compared to the No. 1 Wujiwan， No. 2 significantly reduced SP expression， and the intensity and range of SS expression in the colon tissue in the No. 2 groups were smaller than those in the No. 1 groups （<italic>P</italic><0.05）. Conclusion:Wujiwan at different compatibilities was capable of improving gastrointestinal hormone disorder of IBS to reduce intestinal sensitivity. In terms of systemic effect， No. 1 was superior to No. 2， while in terms of local effect， No. 2 was advantageous. No. 1 Wujiwan was superior to No. 2 in the effect on intestinal dynamics， while No. 2 had an advantageous effect on intestinal sensation over No. 1.

Removal of transepiphyseal plate after fixation for a certain period of time: Growth inhibition of epiphyseal plate after a period of observation / 中国组织工程研究

BACKGROUND: Clinically, children with metaphyseal and periepiphyseal fractures are more common. Internal fixation of the tarsal plate has a more important role in stabilizing the fracture. However, after a certain period of internal fixation, the fixator was taken out. After a certain period of observation, the recovery of the epiphyseal plate was rarely reported. OBJECTIVE: To design an animal model of epiphyseal plate fracture and observe the growth and inhibition of epiphyseal plate after removal of the transepiphyseal steel plate implanted with locking plate for a period of time. METHODS: The 5 mm fracture models of the distal epiphyseal plate of the right femur in 32 young rabbits were established. They were randomly divided into four groups with eight rats in each group. The same type of steel plate and screw was used. Internal fixation of transepiphyseal plate was conducted at periplate fracture line. The internal fixator was removed 2, 4, 8, and 12 weeks after operation. The rabbits were sacrificed after 2 weeks of observation. The femoral specimens were obtained, and the femoral length was measured. The thickness of epiphyseal plate and the number of mast cells were measured by pathological section. Morphological changes of mast cells and epiphyseal plate thickness were observed. The fracture model was used as the experimental group and the distal epiphyseal plate of the left femur was used as the control group. RESULTS AND CONCLUSION: (1) After 2 weeks of internal fixation, the steel plate was removed in the experimental group and the observation was continued for 2 weeks. There was no significant difference in femoral length, epiphyseal thickness and mast cell count between the experimental group and the control group. (2) In the experimental group, the internal fixator was removed at 4, 8 and 12 weeks and the plate was observed for 2 weeks. Compared with the control group, the femoral length, epiphyseal thickness and mast cell count in the experimental group were not completely restored to normal, and the difference was significant (P < 0.05). (3) On the premise that the internal fixator did not injure the epiphyseal plate, the transepiphyseal plate was taken out at the initial stage of internal fixation (≤ 2 weeks), and the plate was observed for 2 additional weeks. The growth and development of the epiphyseal plate were not significantly affected by appropriate pressure. (4) If the pressure limitation lasts for too long (≥ 4 weeks), the time of internal fixation for epiphyseal plate pressure limitation is too long. Although the plate is removed in time, the indexes such as limb length, epiphyseal plate thickness and mast cell count cannot be completely restored to normal, which can lead to partial or complete blockade of epiphyseal plate growth, resulting in limb deformity and stagnation of epiphyseal plate development.

Electroacupuncture improves cutaneous allergic reaction by inhibiting degranulation of intrape-ritoneal mast cells, MAPK signaling and inflammatory factor levels in urticaria rats / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) on degranulation of intraperitoneal mast cells (MCs) and expression of mitogen-activated protein kinase (MAPK) signaling related proteins, tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in urticaria rats, so as to reveal its mechanisms underlying improvement of urticaria. METHODS: Thirty-two SD rats were randomly divided into control，model，EA and medication groups (n＝8 in each group). The urticaria model was established by using passive cutaneous anaphylaxis (PCA) reaction method. EA (2 Hz /15 Hz, 1 mA) was applied to bilateral "Zusanli"(ST36), "Quchi "(LI11) and "Xuehai"(SP10) for 20 min，once daily for 7 consecutive days before antigen attack. Rats of the medication group were treated by gavage of Loratadine(1 mg•kg－1•d－1)for 7 days. The diameter of cutaneous Evan's blue spots was measured to evaluate the severity of PCA. Intraperitoneal fluid smears were prepared to observe the degranulation state of MCs. The contents of TNF-α and IL-6 in the intraperitoneal fluid were detected by ELISA, and the expression of extracellular signal-regulated kinase (ERK), phosphorylated (p)-ERK, c-Jun N-terminal kinase (JNK), p-JNK, P38MAPK and p-P38MAPK of the acquired intraperitoneal MCs was detected by Western blot. RESULTS: The diameter of cutaneous Evan's blue spot was significantly increased in the model group than that in the control group (P<0.01), and considerably decreased in both EA and medication groups compared with the model group(P<0.01). After modeling，the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of ERK, p-ERK, JNK, p-JNK, P38MAPK and p-P38MAPK were remarkably increased in the mo-del group than those in the control group (P<0.01, P<0.05). After the treatment, the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of p-ERK, JNK, p-JNK and p-P38MAPK were obviously decreased in both EA and medication groups relevant to the model group (P<0.01, P<0.05), while no significant changes were found in the expression of ERK in both EA and medication groups, and P38MAPK in the EA group. Compared with the model and EA groups, expression levels of P38MAPK were down-regulated in the medication group (P<0.05). CONCLUSION: EA can reduce skin allergic reaction in rats with urticaria, which may be related to its effects in inhibiting the degranulation of intraperitoneal MCs, down-regulating the expression of MAPK signaling-related proteins and the level of pro-inflammatory factors TNF-α and IL-6 in intraperitoneal MCs.

Effect of electroacupuncture on expressions of Lyn and Syk in mast cells of subcutaneous loose connective tissue in rats with urticarial / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) preconditioning on the expressions of tyrosine kinase Lyn and spleen tyrosine kinase (Syk) in mast cells of subcutaneous loose connective tissue in the rats with urticaria and explore the potential biological mechanism of EA in the intervention of urticaria.@*METHODS@#A total of 32 SD rats were randomized into a blank group, a model group, an EA group and a positive medication group, 8 rats in each one. Except of the blank group, the passive cutaneous anaphylaxis (PCA) was adopted to prepare the model of urticaria in the rats of the rest three groups. In the EA group, EA was applied to bilateral "Quchi" (LI 11), "Xuehai" (SP 10) and "Zusanli" (ST 36), with disperse-dense wave, 2 Hz/15 Hz in frequency and 1 mA in current intensity, once daily, for 20 min each time, consecutively for 7 days. In the positive medication group, loratadine (1 mg•kg•d) was for intragastric administration, once daily, consecutively for 7 days. The samples were collected for index detection 30 min after PCA antigen challenge in the rats of each group. Spectrophotometer was adopted to determine the effusion quantity of Evans blue in the allergized site of skin. HE staining was used to observe the morphological changes in the allergized site of skin. Toluidine blue staining was provided to observe mast cell degranulation in subcutaneous loose connective tissue in the allergized site of skin. Immunohistochemistry was applied to determine the protein expressions of Lyn and Syk during degranulation of mast cells.@*RESULTS@#In the rats of the odel group, the eipdermis of allergized site was thickening, cells were disorganized in hierarchy and inflammatory cells were infiltrated largely in the dermis. In the positive medication group and the EA group, the epidermis was getting thin, cell arrangement was clear and the inflammatory cell infiltration was obviously alleviated as compared with the model group. Compared with the blank group, the OD value of skin dye effusion quantity, the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all increased in the model group (<0.01). Compared with the model group, the OD value of skin dye effusion quantity, the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all reduced in the EA group and the positive medication group (<0.01). Compared with the positive medication group, the degranulation rate of mast cells was increased significantly in the EA group (<0.01).@*CONCLUSION@#Electroacupuncture at "Quchi" (LI 11), "Xuehai" (SP 10) and "Zusanli" (ST 36) reduces vascular permeability and gives play to the role of anti-allergy by the way of regulating and controlling the degranulation of mast cells in the rats with urticaria and the effect mechanism of electroacupuncture may be related to the inhibition of protein expressions of Lyn and Syk in mast cells.

Bullous pemphigoid with neurofibroma-like histopathological change in two patients without neurofibromatosis type 1: Coincidence or association?

@#The author reports two cases of Bullous pemphigoid (BP) with neurofibroma (NF)-like histopathological change. The two patients without neurofibromatosis type 1 (NF1) presented with several bullae on their trunk. Based on the results of positivity for anti-BP180 antibody, direct immunofluorescence, and histopathological findings, they were diagnosed with BP. Histologically, another lesion in the dermis, which was composed of spindle cells with wavy nuclei, collagen fibers, and mast cells, was located close to the bulla. Immunohistochemically, the spindle cells were diffusely positive for S-100 protein and CD34, and weakly positive for epithelial membrane antigen in certain foci. These findings were considered to be “NF-like” histopathological change. This is the first two cases of BP with NF-like histopathological change in patients without NF1.

Mechanism of Yupingfeng Granule on CU Rats Based on IL-23/IL-17 Inflammation Axis / 中国实验方剂学杂志

Objective::To study the effect of Yupingfeng granule on the degranulation of skin mast cells in chronic urticaria (CU) rats and the intervention mechanism of interleukin-23(IL-23), interleukin-17(IL-17) inflammation axis. Method::Totally 60 SPF SD rats were selected and randomly divided into normal group (normal saline), model group (normal saline), and loratadine group (0.9 mg·kg-1·d-1), high-dose Yupingfeng granules group (4.05 g·kg-1·d-1), middle-dose group (2.7 g·kg-1·d-1), low-dose group (1.35 g·kg-1·d-1). The CU rat model was reproduced through intraperitoneal injection of ovalbumin with aluminum hydroxide suspension and DTP vaccine. Histopathological changes of rat skin were observed by hematoxylin-eosin (HE) staining. Degranulation of mast cells in rat skin was determined by toluidine blue staining. IL-23 and IL-17 protein expressions in skin tissue were determined by immunohistochemistry (IHC). IL-23 and IL-17 mRNA transcription levels in skin tissue were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result::Yupingfeng granules can significantly alleviate the pathological manifestations of dermal edema, collagen beam distance, inflammatory cell infiltration of CU rats, and reduce the degranulation reaction of skin tissue mast cells in CU rats. The IL-23, IL-17 mRNA and protein expressions of the skin of model group were significantly increased compared with the normal group (P<0.05, P<0.01). Compared with the model group, Yupingfeng granules can significantly down-regulate IL-23 mRNA and protein expressions of CU rats (P<0.05, P<0.01). Yupingfeng granules had no significant regulatory effect on IL-17. Conclusion::Yupingfeng granule can significantly reduce the degranulation of mast cells in skin tissue of CU rats, and improve the pathological manifestations, such as dermal edema, serous exudation and inflammatory cell infiltration. The mechanism may be related to inhibiting the secretion of IL-23 pro-inflammatory cytokines and improving CU lesions.

Clinical, Laboratory, and Bone Marrow Findings of 31 Patients With Waldenström Macroglobulinemia

BACKGROUND: Waldenström macroglobulinemia (WM) is a subset of lymphoplasmacytic lymphoma (LPL) with bone marrow (BM) involvement and an IgM monoclonal gammopathy of any level. We aimed to identify the clinical, laboratory, and BM findings of patients with WM and to evaluate the usefulness of CD154 for the diagnosis and prognosis of WM.METHODS: We reviewed the medical records and BM studies and/or flow cytometric immunotyping of 31 patients with untreated WM. Semiquantitative immunohistochemistry (CD20, CD138, tryptase, and CD154) of BM was performed.RESULTS: Only six patients presented with symptoms of hyperviscosity syndrome. Eleven patients had solid cancer and/or another hematologic malignancy. Mast cells (MC) increased in all samples, with some in close contact with tumor cells. Tryptase-positive MC (17.1/ high-power fields [HPF], 1.2–72.0/HPF) and CD154-positive MC (8.6/HPF, 0.1–31.1/HPF) were observed. The high CD154-positive MC (≥8.6/HPF) group showed a lower overall five-year survival rate than the low CD154-positive MC (<8.6/HPF) group (71.9% vs. 100.0%; P=0.012). Flow cytometric immunophenotyping of BM aspirates showed increased B lymphocytes and plasma cells with a normal phenotype (CD138⁺/CD38⁺/CD19⁺/CD45⁺/CD56⁻).CONCLUSIONS: Approximately one third of WM patients showed other malignancies and all patients had increased MC. Immunohistochemistry and flow cytometric immunophenotyping are useful for diagnosing WM, and increased CD154-positive MC can indicate poor prognosis.

Uma visão atualizada da anafilaxia idiopática: a ausência de evidência não é a evidência de ausência / An updated view of idiopathic anaphylaxis: absence of evidence is not evidence of absence

A anafilaxia idiopática não apresenta etiologia conhecida. A sua prevalência é estimada entre 10-35% de todas as modalidades de anafilaxia. A sintomatologia apresentada é a mesma de qualquer outra anafilaxia: urticária, angioedema, ruborização, prurido, hipotensão arterial, taquicardia, manifestações gastrointestinais (disfagia, náusea, vômitos, cólicas abdominais, diarreia), asma, edema laríngeo, tontura e síncope. A mortalidade é rara. Não há transmissão genética, mas 40% dos pacientes são atópicos. É mais frequente nos adultos do que nas crianças, e principalmente em mulheres. É um diagnóstico de exclusão. Ocorre ativação mastocitária com desgranulação citoplasmática dos mediadores de anafilaxia (triptase, histamina, entre outros). É uma anafilaxia com boa resposta aos corticoides, e, portanto, caso não haja resposta adequada a doses eficazes de prednisona/prednisolona, o seu diagnóstico deve ser revisto. O diagnóstico diferencial da anafilaxia idiopática inclui: a mastocitose sistêmica indolente, síndromes de ativação mastocitária monoclonais, alergia à galactose-alfa-1,3 galactose, anafilaxia induzida por exercícios (com e sem dependência alimentar e medicamentosa), angioedema hereditário (congênito e adquirido), feocromocitoma, síndrome carcinoide, anafilaxia oral acarina, alergia ao Anisakis simplex, disfunção das cordas vocais, síndrome escombroide, alergia ao sêmen, alergia ao látex, manifestações psicossomáticas (síndrome do pânico, globus hystericus e a síndrome de Münchausen), bem como as tradicionais e mais frequentes modalidades de anafilaxia (alergia a alimentos, medicamentos e insetos). O tratamento na crise aguda da anafilaxia idiopática é o mesmo do que nas demais anafilaxias, incluindo a administração intramuscular imediata de epinefrina. Deve haver uma generosa e prolongada prescrição de corticoterapia oral, e também a instituição de medicação preventiva (anti-histamínicos anti- H1 e anti-H2, cetotifeno, albuterol oral, montelucaste, cromoglicato de sódio, e por último o omalizumabe). Os pacientes devem portar epinefrina autoinjetora e ser instruídos sobre como agir em caso de um episódio anafilático. Eles respondem bem à administração de epinefrina. A corticoterapia oral, por 4-6 semanas, pode induzir uma remissão completa.

Idiopathic anaphylaxis is a condition of unknown etiology. Its prevalence ranges from 10 to 35% of all cases of anaphylaxis. Clinical symptoms and signs are those of classic anaphylaxis, including urticaria, angioedema, flushing, itching, hypotension, tachycardia, gastrointestinal manifestations (dysphagia, nausea, vomiting, abdominal cramps, and diarrhea), asthma, laryngeal edema, dizziness, and syncope. Mortality is rare. There is no genetic transmission, but about 40% of patients are atopic. It is more common in adults than in children, affecting mainly women. It is considered a diagnosis of exclusion of other known forms of anaphylaxis. Mast cell activation occurs with cytoplasmatic degranulation of mediators of anaphylaxis (tryptase and histamine, among others). Because idiopathic anaphylaxis is a steroid-responsive condition, if it is not controlled with adequate doses of prednisone/prednisolone, the diagnosis should be challenged. The differential diagnosis of idiopathic anaphylaxis includes indolent systemic mastocytosis, clonal mast cell activation syndromes, galactose-alpha-1,3- galactose allergy, exercise-induced anaphylaxis (both food- and drug-dependent and -independent), hereditary angioedema (congenital and acquired), pheochromocytoma, carcinoid syndrome, oral mite anaphylaxis, Anisakis simplex allergy, vocal cord dysfunction, scombroid poisoning, semen allergy, latex allergy, psychosomatic conditions (panic attacks, globus hystericus, and Münchausen syndrome), and the classic forms of anaphylaxis (food, drug, and insect allergies). Treatment of acute idiopathic anaphylaxis is the same as in the other forms of anaphylaxis, including intramuscular epinephrine, but with prolonged oral corticosteroid therapy. It might also include other oral preventive medications (H1 and H2 antihistamines, ketotifen, oral albuterol, montelukast, sodium cromoglycate, and recently omalizumab). Patients should have an epinephrine auto-injector and be instructed on self-management of anaphylaxis. Good response to epinephrine is observed, and oral corticosteroid therapy for 4-6 weeks can induce complete remission.

Comparative Evaluation of Efficacy and Safety of BepotastineBesilate versus Olopatadine and Ketorolac Combination in Patients with Vernal Keratoconjunctivitis

Objective: Comparative evaluation of efficacy and safety ofBepotastine besilate versus Olopatadine and Ketorolaccombination in patients with vernal keratoconjunctivitis.Materials and Methods: This was a prospective, open label,randomized, comparative clinical study. Hundred patients ofvernal keratoconjunctivitis between 6 to 20 years of age ofeither sex willing to give informed consent were enrolled in thestudy. In Group 1, 50 patients received Bepotastine besilate(0.15%) eye drops twice daily for 8 weeks whereas in Group 2,50 patients received Olopatadine (0.2%) and Ketorolac (0.4%)combination eye drops twice daily for 8 weeks. Symptoms andsigns scoring of VKC were recorded on baseline and at thetime of follow up at 4 and 8 weeks. Safety assessments werealso done in both the drug groups during the study period forany serious adverse effects.Results: After the 2 months of drug therapy, patients in boththe groups showed improvement in the symptoms and signsscoring of VKC. However, there was no statistically significantdifference between the two treatment groups at 4th and 8thweek. Both the drugs were well tolerated without any seriousadverse effect.Conclusion: Both bepotastine besilate versus olopatadine andKetorolac combination ophthalmic solutions were found to beeffective in alleviating the clinical symptoms and signs of VKC.

The effects of mobile phone exposure on mast cells in rat dura mater / Efectos de la exposición del teléfono móvil en los mastocitos de la duramadre de ratas

Mobile phone use has increased rapidly. The central nervous system has been shown to be adversely affected by its electromagnetic field (EMF) resulting in headache and sleep disturbances. How the cells make up the CNS and are affected by EMF is unclear. However, because of their central role in inflammation through diverse stimuli including radiation, this study aimed to investigate the effects of electromagnetic fields induced by mobile phones on mast cells in rat dura mater. A total of 18 adult, female, SpragueDawley rats were divided into two groups. The choice of female rats for his study was based on recent surveys demonstrating that mobile phone use is more frequent and prolonged among females. The study group was exposed to 900 MHz electromagnetic field (1 h/day for 45 days). In the end of the study, duramater tissue was extracted and stained using Toluidine blue. Mast cells were counted and results were analysed using Student t test. Mean mast cell number was 202.33±9.82 and 456.78±35.01 in the control and study groups, respectively (p<0.05). Analysis of serum electrolyte and immunoglobulin E levels showed no statistically significant difference between the two groups (p>0.05). The study showed that mobile phone exposure increased mast cell number and degranulation in rat dura mater. Further studies are required to evaluate the clinical implications of these findings.

El uso del teléfono móvil ha aumentado rápidamente. Se ha demostrado que el sistema nervioso central (SNC) se ve afectado de manera adversa debido al campo electromagnético (CEM) que produce dolor de cabeza y trastornos del sueño. No está claro cómo se ve afectada la composición celular del SNC por el CEM. Sin embargo, debido a su función principal en la inflamación a través de diversos estímulos que incluyen la radiación, este estudio tuvo como objetivo investigar los efectos de los campos electromagnéticos inducidos por los teléfonos móviles en los mastocitos de la duramadre de ratas. Un total de 18 ratas Sprague-Dawley adultas, hembras, se dividieron en dos grupos. Se usaron ratas hembras para este estudio en base a investigaciones recientes que han demostrado que el uso de teléfonos móviles es más frecuente y prolongado en las mujeres. Los grupos de estudio fueron expuestos a un campo electromagnético de 900 MHz (1 h / día durante 45 días). Al término del estudio, fue extirpado el tejido de la duramadre y teñido con azul de toluidina. Se contaron los mastocitos y se analizaron los resultados utilizando la prueba t de Student. La cantidad media de células cebadas fue de 202,33 ± 9.82 y 456,78 ± 35,01 en los grupos control y estudio, respectivamente (p <0,05). El análisis del electrolito sérico y los niveles de inmunoglobulina E no mostraron diferencias estadísticamente significativas entre los dos grupos (p> 0,05). El estudio mostró que la exposición a teléfonos móviles aumentó el número de mastocitos y la desgranulación en la duramadre de las ratas. Se requieren estudios adicionales para evaluar las implicaciones clínicas de estos hallazgos.

Use of different fixation times and application of two immunohistochemical methods for detection of KIT and Ki67 proteins in canine cutaneous mast cell tumors / Utilização de diferentes tempos de fixação e métodos imuno-histoquímicos na detecção das proteínas KIT e Ki67 em mastocitomas cutâneos caninos

Due to the high prevalence of mast cell tumors (MCTs) in the diagnostic routine, several factors, especially prognostic, have been sought to determine the biological behavior of these neoplasms. Immunohistochemistry (IHC) is one of the main tools utilized to biologically differentiate more aggressive tumors from less aggressive ones. However, some immunostainings are influenced by formalin fixation, interfering with the results. This is both a retrospective and prospective study of MCTs diagnosed in laboratory routine. A total of 25 samples, without knowledge about fixation time, were analyzed in the retrospective study, whereas 12 samples, with known fixation times, were assessed in the prospective study. Two histologic grading systems (Patnaik and Kiupel), special staining of toluidine blue, and IHC for KIT and Ki67 proteins were applied in both studies. Additionally, two amplification systems (biotinylated and non-biotinylated) for Ki67 protein and counting of the argyrophilic nucleolar organizing regions (AgNOR method) were tested in the prospective study. In the retrospective study, greater agreement between the evaluating pathologists was observed when the Kiupel system was used. IHC staining for KIT protein was effective in both studies, regardless of fixation time. IHC staining for Ki67 protein was highly sensitive to formaldehyde, and staining failure was observed in 56% of the cases in the retrospective study. In the prospective study, samples fixed for longer than 24 hours showed a reduction in the number of stained cells (altering the determination of the cell growth fraction) or showed absence of IHC staining in both amplification systems. The use of the AgNOR method to evaluate the rate of cell proliferation may be an alternative when the fixation time of the neoplasm is unknown or longer than 24 hours.(AU)

Devido a alta prevalência dos mastocitomas cutâneos caninos (MCCs) na rotina diagnóstica, vários fatores, especialmente fatores prognósticos, têm sido buscados para auxiliar na determinação do comportamento biológico desse neoplasma. A imuno-histoquímica é uma das principais ferramentas empregadas para diferenciar tumores biologicamente mais agressivos de tumores menos agressivos. Entretanto, algumas imunomarcações sofrem influência pela fixação em formol, interferindo nos resultados. Este estudo compreendeu avaliar através de uma etapa retrospectiva e uma etapa prospectiva casos de MCCs diagnosticados na rotina laboratorial. Um total de 25 amostras, sem conhecimento do tempo de fixação, foi analisado no estudo retrospectivo e 12 amostras, com tempos de fixação conhecidos, no estudo prospectivo. Foram aplicados nos dois estudos, dois sistemas de graduação histológica (Patnaik e Kiupel), a coloração especial de azul de toluidina e a imuno-histoquímica para as proteínas KIT e Ki67. Adicionalmente, no estudo prospectivo, foram testados dois sistemas de amplificação (biotinilado e não biotinilado) para a proteína Ki67 e a técnica de AgNOR (contagem das regiões organizadoras nucleolares argirofílicas). Na etapa retrospectiva, observou-se uma maior concordância entre os patologistas avaliadores quando o sistema Kiupel foi utilizado. A imunomarcação para KIT se manteve eficaz em ambos os estudos, independentemente do tempo de fixação. A imunomarcação para o Ki67 mostrou-se altamente sensível ao tempo de fixação em formol, sendo observada falha na imunomarcação em 56% dos casos do estudo retrospectivo. No estudo prospectivo, constatou-se que amostras fixadas por mais de 24 horas em formol apresentaram redução na quantidade de células imunomarcadas (alterando a determinação da fração de crescimento celular) ou apresentaram ausência de imunomarcação em ambos os sistemas de amplificação. A utilização do método AgNOR, para avaliar a taxa de proliferação celular, pode ser uma alternativa quando o tempo de fixação do neoplasma for desconhecido ou superior a 24 horas.(AU)

Role of hippocampal mast cells in early postoperative cognitive impairment in rats / 中华麻醉学杂志

Objective@#To evaluate the role of hippocampal mast cells in the early postoperative cognitive impairment in rats.@*Methods@#Eighty male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 4 groups (n=20 each) using a random number table method: normal saline control group (group C), cromolyn injected into lateral cerebral ventricle group (group L), operation group (group O), and cromolyn injected into lateral cerebral ventricle plus operation group (group LO). Cromolyn 2 μl (100 μg/ul) was intracerebroventricularly injected in L and LO groups.The equal volume of normal saline was given instead in C and O groups.Open reduction and internal fixation was performed after tibial fracture was induced 30 min later in O and LO groups.Contextual fear conditioning and Y-maze tests were performed at 1 and 3 days after operation, and the freezing time and the number of learning trails were recorded.The animals were then sacrificed, and hippocampal tissues were obtained for determination of activated mast cell count (by toluidine blue staining) and expression of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) mRNA (by polymerase chain reaction).@*Results@#Compared with group C, the number of learning trails was significantly increased, and the freezing time was shortened, the number of activated mast cells in hippocampi was increased, and the expression of IL-1β mRNA was up-regulated at 1 and 3 days after operation, and the expression of TNF-α mRNA was up-regulated at 1 day after operation in group O(P<0.05), and no significant change was found in the parameters mentioned above in group L (P>0.05). Compared with group O, the number of learning trails was significantly decreased, and the freezing time was prolonged, the number of activated mast cells in hippocampi was decreased, and the expression of IL-1β mRNA in hippocampi was down-regulated at 1 and 3 days after operation, and the expression of TNF-α mRNA in hippocampi was down-regulated at 1 day after operation in group LO (P<0.05).@*Conclusion@#Activation of hippocampal mast cells can induce central inflammatory responses and is involved in the mechanism of early postoperative cognitive impairment in rats.