Acupuncture Alleviates Menstrual Pain in Rat Model via Suppressing Eotaxin/CCR3 Axis to Weak EOS-MC Activation

Introduction. Emerging data show that chemokine-mediated inflammation is involved in the occurrence and maintenance of pain. Recent evidence suggests that eotaxin levels rise when dysmenorrhea happens. The purpose of this study is to investigate whether eotaxin/CC chemokine receptor 3 (CCR3) axis, a key regulatory pathway for eosinophils (EOS) recruitment, is involved in acupuncture analgesia for dysmenorrhea. Methods. After the cold congealing dysmenorrhea (CCD) rat model prepared, animals received perpendicular needling (PN) and transverse needling (TN) at SP6, respectively, for 20 min. The CCR3 agonist CCL11 was administered 30 min prior to acupuncture. Pain behavior was assessed via a writhing response. The uterine contraction test was detected by an electrophysiological method. Eotaxin, histamine (HIS), and interleukin-6 (IL-6) levels were evaluated by ELISA. The expression of CCR3 and histamine H1 receptor (H1R) was analyzed by RT-qPCR and Western blot. The expression of EOS, mast cells (MCs), eosinophil peroxidase (EPO), and eosinophil cationic protein (ECP) was assessed by hematoxylin-eosin staining (HE), Toluidine Blue staining (TB), and immunohistochemistry, respectively. Results. Acupuncture prominently attenuated the menstrual pain in CCD rats, particularly TN technique. Electrophysiological recording data showed that the increased uterine contractility was ameliorated by acupuncture. In addition, TN decreased the release of eotaxin, HIS, IL-6, and the expression of CCR3 and H1R. HE, TB staining, and immunohistochemistry experiments showed that the increased expression of EOS, MCs, EPO, and ECP in uterine tissues was reversed by TN. Furthermore, we found that the effects of TN against CCD-induced menstrual pain, increased ECP expression, and HIS level were abolished by CCL11. Conclusion. TN alleviated menstrual pain by improving the uterine inflammatory environment via suppressing eotaxin/CCR3 axis to weak EOS-MC activation in CCD rats. The study findings support the acupuncture as a promising approach for dysmenorrhea, meanwhile, indicating the importance of performing appropriate needling technique.

IL-37 expression improves renal function after ischemia and reperfusion in mice model

Renal ischemia is a major problem in the world that lead to renal failure for which no effective treatment is available. Renal ischemia involves a robust inflammatory response, involving up-regulated chemokine expression and leukocyte accumulation, contributes to the mechanism of renal injury and renal failure. IL-37 is a new human cytokine and has an anti-inflammatory function. Currently, it is unknown whether IL-37 suppresses renal inflammatory response to ischemia. We tested the hypothesis that expression of human IL-37 in mouse protects the renal against ischemic injury through suppression of the renal inflammatory response. IL-37 Tg and WT mice were subjected to right renal nephrectomy to induce unilateral model of ischemia the microvascular clamp was positioned around the left renal pedicles. Serum sampling for measurements of TNF-α, IL-1β, Caspase3, MDA, HMGB1, urea and creatinine. Hematoxylin-eosin staining for histological analysis. The resulted data showed that IL-37 has anti-inflammatory effects in renal IRI as evidenced by significant reduction of the inflammatory markers levels TNF-α, IL-1β and HMGB1. IL-37 has potent antioxidant and anti-apoptotic effects with significant reduction in MDA and caspace-3 respectively

IL-37 expression improves renal function after ischemia and reperfusion in mice model

Renal ischemia is a major problem in the world that lead to renal failure for which no effective treatment is available. Renal ischemia involves a robust inflammatory response, involving up-regulated chemokine expression and leukocyte accumulation, contributes to the mechanism of renal injury and renal failure. IL-37 is a new human cytokine and has an anti-inflammatory function. Currently, it is unknown whether IL-37 suppresses renal inflammatory response to ischemia. We tested the hypothesis that expression of human IL-37 in mouse protects the renal against ischemic injury through suppression of the renal inflammatory response. IL-37 Tg and WT mice were subjected to right renal nephrectomy to induce unilateral model of ischemia the microvascular clamp was positioned around the left renal pedicles. Serum sampling for measurements of TNF-α, IL-1β, Caspase3, MDA, HMGB1, urea and creatinine. Hematoxylin-eosin staining for histological analysis. The resulted data showed that IL-37 has anti-inflammatory effects in renal IRI as evidenced by significant reduction of the inflammatory markers levels TNF-α, IL-1β and HMGB1. IL-37 has potent antioxidant and anti-apoptotic effects with significant reduction in MDA and caspace-3 respectively. Keywords: Renal ischemia, IL-37, TNF-α, IL-1β, Caspase-3

IL-37 expression improves renal function after ischemia and reperfusion in mice model

Renal ischemia is a major problem in the world that lead to renal failure for which no effective treatment is available. Renal ischemia involves a robust inflammatory response, involving up-regulated chemokine expression and leukocyte accumulation, contributes to the mechanism of renal injury and renal failure. IL-37 is a new human cytokine and has an anti-inflammatory function. Currently, it is unknown whether IL-37 suppresses renal inflammatory response to ischemia. We tested the hypothesis that expression of human IL-37 in mouse protects the renal against ischemic injury through suppression of the renal inflammatory response. IL-37 Tg and WT mice were subjected to right renal nephrectomy to induce unilateral model of ischemia the microvascular clamp was positioned around the left renal pedicles. Serum sampling for measurements of TNF-α, IL-1β, Caspase3, MDA, HMGB1, urea and creatinine. Hematoxylin-eosin staining for histological analysis. The resulted data showed that IL-37 has anti-inflammatory effects in renal IRI as evidenced by significant reduction of the inflammatory markers levels TNF-α, IL-1β and HMGB1. IL-37 has potent antioxidant and anti-apoptotic effects with significant reduction in MDA and caspace-3 respectively.

Inflammation in rats born to mothers treated with dexamethasone 15cH during pregnancy: an immunohistochemical study

In previous studies, we observed that rats born to mothers treated with dexamethasone 15CH (10-33M) had a higher level of mast cell degranulation and greater arteriolar dilation after the exposure of an inflammatory stimulus, suggesting the possibility of vertical transmission of the effects of ultra-diluted substances between mother and offspring. In this study, a more detailed assessment of the cellular events in acute inflammation was made using techniques of immunohistochemistry. The identification of adhesion molecules expression was made by the markers: anti-CD54 (ICAM-1) and anti-CD18 (β2-Integrin). The identification of inflammatory cells was performed by the markers anti-MAC387 (mononuclear cells) and anti-CD163 (active macrophages). Polymorphonuclear cells were identified by hematoxylin-eosin staining. The number of labeled cells per field was recorded, except for the anti-CD54 marker, whose intensity of staining on the endothelial cells was defined by scores assigned by two independent observers. The results point toward to an up regulation of the whole inflammatory process in rats born to mothers treated with dexamethasone 15CH during pregnancy. This conclusion is justified by the following statistically significant (p≤0.05) findings: a) bigger mast cell degranulation and increased of arteriolar diameter; b) increased migration of polymorphonuclear cells in relation to the mononuclear cells; c) earlier expression of CD163 in monocytes, d) higher level of adhesion molecules expression.

Rasio Penutupan Luka pada Tikus Diabetes Diinduksi Streptozotocin dengan Perlakuan Dressing Tipe Pasif dan Interaktif (Penelitian Pendahuluan)

Deparaffinization is a stage before the staining process to remove/dissolve paraffin so that the absorption of color in tissue preparations is maximized. Deparaffinization is usually carried out using xylol and toluol. Xylol has toxic effects including acute neurotoxicity, heart and kidney damage, hepatotoxicity, fatal blood dyscrasias, skin erythema, dry skin, peeling skin, and also has a carcinogenic effect. The toxicity effect of olive oil is lower than that of xylol. Oils that have non-polar properties can remove the remaining paraffin contained in the tissue. The purpose of this study was to determine the microscopic appearance of the kidney tissue preparations of mice deparaffinized with olive oil on hematoxylin eosin (HE) staining. The type of research used is experimental research which is analyzed with a descriptive approach. The results of the assessment of preparations deparaffinized with xylol in 80 visual fields obtained 100% good preparations and preparations deparaffinized with olive oil in 80 visual fields obtained 0% poor preparations, 11.3% poor preparations, and 88.7% good preparation. So it can be said that better results are found in the microscopic picture of the kidney preparations of mice (Mus musculus) deparaffinized with xylol.

Efek pemberian α-tokoferol terhadap jumlah sel spermatogenik dan sel Leydig pada tikus putih (Rattus norvegicus) yang dipapar 2,3,7,8 Tetrachlorodibenzo-p-dioxin

This research aimed to determine the effect of α-tocopherol on the count of spermatogenic and Leydig cells in rats (Rattus norvegicus) exposed to 2,3,7,8 tetrachlorodibenzo-p-dioxin (dioxin). Dioxin is an endocrine-disrupting agent that adversely affects reproductive health, while α-tocopherol maintains fertility. This research used 25 rats aged 10-12 weeks weighing 150-200 grams. Rats were divided into five groups (K, P0, P1, P2 and P3). The K (control group) was administered with corn oil 1 ml/day. P0 was exposed to 700 ng/kg/day dioxin. P1, P2 and P3 was exposed to dioxin at a dose of 700 ng/kg/day and administered with α-tocopherol at a dose of 77, 140 and 259 mg/kg/day respectively. Dioxin exposure, corn oil and α-tocopherol administration were conducted orally for 20 days. On day-21, all rats were sacrificed for histological slides preparation of testicles with hematoxylin-eosin staining. Data were analyzed with analysis of variance and continued with the Duncan test. The results indicated that exposure to dioxin caused a decrease in the number of spermatogenic and Leydig cells. The administration of α-tocopherol at a dose of 140 mg/kg/day eliminated the effect of reducing the number of spermatogenic and Leydig cells caused by exposure to dioxin. The conclusion was the administration of α-tocopherol at 140 mg/kg/day was effective in maintaining the number of spermatogenic and Leydig cells in rats (Rattus norvegicus) exposed to 2,3,7,8 tetrachlorodibenzo-p-dioxin.

Long-Term Evaluation of Poly(lactic acid) (PLA) Implants in a Horse: An Experimental Pilot Study

In horses, there is an increasing interest in developing long-lasting drug formulations, with biopolymers as viable carrier alternatives in addition to their use as scaffolds, suture threads, screws, pins, and plates for orthopedic surgeries. This communication focuses on the prolonged biocompatibility and biodegradation of PLA, prepared by hot pressing at 180 °C. Six samples were implanted subcutaneously on the lateral surface of the neck of one horse. The polymers remained implanted for 24 to 57 weeks. Physical examination, plasma fibrinogen, and the mechanical nociceptive threshold (MNT) were performed. After 24, 28, 34, 38, and 57 weeks, the materials were removed for histochemical analysis using hematoxylin-eosin and scanning electron microscopy (SEM). There were no essential clinical changes. MNT decreased after the implantation procedure, returning to normal after 48 h. A foreign body response was observed by histopathologic evaluation up to 38 weeks. At 57 weeks, no polymer or fibrotic capsules were identified. SEM showed surface roughness suggesting a biodegradation process, with an increase in the median pore diameter. As in the histopathological evaluation, it was not possible to detect the polymer 57 weeks after implantation. PLA showed biocompatible degradation and these findings may contribute to future research in the biomedical area.

Chrysosplenol D protects mice against LPS-induced acute lung injury by inhibiting oxidative stress, inflammation, and apoptosis via TLR4-MAPKs/NF-κB signaling pathways

This study investigated the effect and mechanism of chrysosplenol D (CD) on LPS-induced acute lung injury in mice. Histological changes in the lungs were measured by hematoxylin-eosin staining. The levels of IL-6, IL-1β, and TNF-α in the bronchoalveolar lavage fluid were detected by ELISA. The levels of oxidative stress were detected by the cuvette assay. Immune cells in peripheral blood, the levels of reactive oxygen species, and apoptosis of primary lung cells were detected by flow cytometry. The mRNA levels of TLR4, MyD88, IL-1β, and NLRP3 were measured by quantitative real-time polymerase chain reaction. The levels of proteins in apoptosis and the TLR4-MAPKs/NF-κB signaling pathways were detected by Western blot. Hematoxylin-eosin staining showed that CD could improve lung injury; decrease the levels of inflammatory factors, oxidative stress, reactive oxygen species, and cell apoptosis; and regulate the immune system. Moreover, CD could down-regulate the mRNA levels of TLR4, MyD88, NLRP3, and IL-1β in lung, and the protein levels of Keap-1, Cleaved-Caspase-3/Caspase-3, Cleaved-Caspase-9/Caspase-9, TLR4, MyD88, p-ERK/ERK, p-JNK/JNK, p-p38/p38, p-p65/p65, NLRP3, and IL-1β, and up-regulated the levels of Bcl-2/Bax, p-Nrf2/Nrf2, and HO-1. The results suggested that CD could protect mice against LPS-induced acute lung injury by inhibiting oxidative stress, inflammation, and apoptosis via the TLR4-MAPKs/NF-κB signaling pathways.

Enteral nutrition ameliorates the symptoms of Crohn's disease in mice via activating special pro-resolving mediators through innate lymphoid cells

Crohn's disease activates the inflammatory reactions to induce intestinal disorders. Enteral nutrition (EN) could exert general immunomodulatory effects. Cecal ligation and perforation (CLP) surgery was utilized to establish Crohn's disease mice models. Survival analysis, hematoxylin-eosin staining, flow cytometry, ELISA, Western blot and liquid chromatography-tandem MS were applied. Baicalein was added to inhibit lipoxygenases. The survival rate was restored and inflammatory injury, exudate neutrophils in peritoneal lavage and serum levels of IL-6 and TNF-α were ameliorated by EN treatment as compared with CLP treatment. EN also increased ILC-3 content, 5/15-LOX level and RvD1-RvD5 in peritoneal lavage. Baicalein reversed all the detected effects of EN except ILC-3 content. EN could activate special pro-resolving mediators (SPMs) through ILCs to mitigate injuries of Crohn's disease.